The Punctum Fixum-Punctum Mobile Model: A Neuromuscular Principle for Efficient Movement Generation?

According to the “punctum fixum–punctum mobile model” that was introduced in prior studies, for generation of the most effective intentional acceleration of a body part the intersegmental neuromuscular onset succession has to spread successively from the rotation axis (punctum fixum) toward the body part that shall be accelerated (punctum mobile). The aim of the present study was to investigate whether this principle is, indeed, fundamental for any kind of efficient rotational accelerations in general, independent of the kind of movements, type of rotational axis, the current body position, or movement direction. Neuromuscular onset succession was captured by surface electromyography of relevant muscles of the anterior and posterior muscle chain in 16 high-level gymnasts during intentional accelerating movement phases while performing 18 different gymnastics elements (in various body positions to forward and backward, performed on high bar, parallel bars, rings and trampoline), as well as during non-sport specific pivot movements around the longitudinal axis. The succession patterns to generate the acceleration phases during these movements were described and statistically evaluated based on the onset time difference between the muscles of the corresponding muscle chain. In all the analyzed movement phases, the results clearly support the hypothesized succession pattern from punctum fixum to punctum mobile. This principle was further underlined by the finding that the succession patterns do change their direction running through the body when the rotational axis (punctum fixum) has been changed (e.g., high bar or rings [hands] vs. floor or trampoline [feet]). The findings improve our understanding of intersegmental neuromuscular coordination patterns to generate intentional movements most efficiently. This could help to develop more specific methods to facilitate such patterns in particular contexts, thus allowing for shorter motor learning procedures of context-specific key movement sequences in different disciplines of sports, as well as during non-sport specific movements.     

Neuromuscular onset succession of high level gymnasts during dynamic leg acceleration phases on high bar

In several athletic disciplines there is evidence that for generating the most effective acceleration of a specific body part the transfer of momentum should run in a “whip-like” consecutive succession of body parts towards the segment which shall be accelerated most effectively (e.g. the arm in throwing disciplines). This study investigated the question how this relates to the succession of neuromuscular activation to induce such “whip like” leg acceleration in sports like gymnastics with changed conditions concerning the body position and momentary rotational axis of movements (e.g. performing giant swings on high bar). The study demonstrates that during different long hang elements, performed by 12 high level gymnasts, the succession of the neuromuscular activation runs primarily from the bar (punctum fixum) towards the legs (punctum mobile). This demonstrates that the frequently used teaching instruction, first to accelerate the legs for a successful realization of such movements, according to a high level kinematic output, is contradictory to the neuromuscular input patterns, being used in high level athletes, realizing these skills with high efficiency.Based on these findings new approaches could be developed for more direct and more adequate teaching methods regarding to an earlier optimization and facilitation of fundamental movement requirements.     

On the organizing role of nonmuscular forces during performance of a giant circle in gymnastics

Five elite gymnasts performed giant circles on the high bar under different conditions of loading (without and with 6-kg loads attached to the shoulders, waist or ankles). Comparing the gymnasts' kinematic pattern of movement with that of a triple-pendulum moving under the sole influence of nonmuscular forces revealed qualitative similarities, including the adoption of an arched position during the downswing and a piked position during the upswing. The structuring role of nonmuscular forces in the organization of movement was further reinforced by the results of an inverse dynamics analysis, assessing the contributions of gravitational, inertial and muscular components to the net joint torques. Adding loads at the level of the shoulders, waist or ankles systematically influenced movement kinematics and net joint torques. However, with the loads attached at the level of the shoulders or waist, the load-induced changes in gravitational and inertial torques provided the required increase in net joint torque, thereby allowing the muscular torques to remain unchanged. With the loads attached at the level of the ankles, this was no longer the case and the gymnasts increased the muscular torques at the shoulder and hip joints. Together, these results demonstrate that expert gymnasts skillfully exploit the operative nonmuscular forces, employing muscle force only in the capacity of complementary forces needed to perform the task.     

The margin for error when releasing the asymmetric bars for dismounts

It has previously been shown that male gymnasts using the "scooped" giant circling technique were able to flatten the path followed by their mass center, resulting in a larger margin for error when releasing the high bar (Hiley and Yeadon, 2003a). The circling technique prior to performing double layout somersault dismounts from the asymmetric bars in women's artistic gymnastics appears to be similar to the "traditional" technique used by some male gymnasts on the high bar. It was speculated that as a result the female gymnasts would have margins for error similar to those of male gymnasts who use the traditional technique. However, it is unclear how the technique of the female gymnasts is affected by the need to avoid the lower bar. A 4-segment planar simulation model of the gymnast and upper bar was used to determine the margins for error when releasing the bar for 9 double layout somersault dismounts at the Sydney 2000 Olympics. The elastic properties of the gymnast and bar were modeled using damped linear springs. Model parameters, primarily the inertia and spring parameters, were optimized to obtain a close match between simulated and actual performances in terms of rotation angle (1.2 degrees), bar displacement (0.011 m), and release velocities (<1%). Each matching simulation was used to determine the time window around the actual point of release for which the model had appropriate release parameters to complete the dismount successfully. The margins for error of the 9 female gymnasts (release window 43-102 ms) were comparable to those of the 3 male gymnasts using the traditional technique (release window 79-84 ms).     

The influence of bar diameter on neuromuscular strength and activation: inferences from an isometric unilateral bench press

The purpose of this study was to examine the influence of two different bar diameters on neuromuscular activation and strength. The bar diameters used reflected a standard Olympic bar (28 mm (1.1 inch); THIN) and a larger fat bar (51 mm [2 inch]; THICK). Eighteen healthy men (age 25.0 +/- 1 years) were assessed for their maximal voluntary contraction (MVC) during a unilateral isometric bench press exercise with the 2 bar types at 2 different joint angles (angle 1 and angle 2; elbow joint at approximately 45 and 90 degrees , respectively). Additionally, on a separate day, subjects performed three 10-second isometric repetitions at an intensity of 80% MVC using the 2 different bars at angle 1 and angle 2. Electromyographic recordings were collected in the pectoralis major and the muscles of the forearm flexor region at a sampling rate of 1000 Hz during the second day of testing. Analysis of variance was used to examine differences in MVC between bars and also examine between bar differences in electromyographic activity for each muscle group at each joint angle. A significance level of 0.05 was used for all tests. MVC was not different between bar types, although there was a main effect of joint angle on MVC such that it was greater at angle 2. There was a main effect of bar at both angles for the forearm muscles and at angle 1 for the pectoralis such that electromyographic activity was greater with THIN. Our data do not support the hypothesis that bar diameter influences performance during an isometric bench press exercise. However, higher electromyographic activity with THIN suggests greater neuromuscular activation with a standard Olympic bar as opposed to a larger diameter "fat" bar. Although our data do not support the use of a fat bar for increasing neuromuscular activation, these findings should be confirmed in other resistance training exercises.     

Age-related changes in neuromuscular function of the quadriceps muscle in physically active adults

Substantial evidence exists for the age-related decline in maximal strength and strength development. Despite the importance of knee extensor strength for physical function and mobility in the elderly, studies focusing on the underlying neuromuscular mechanisms of the quadriceps muscle weakness are limited.The aim of this study was to investigate the contributions of age-related neural and muscular changes in the quadriceps muscle to decreases in isometric maximal voluntary torque (iMVT) and explosive voluntary strength. The interpolated twitch technique and normalized surface electromyography (EMG) signal during iMVT were analyzed to assess changes in neural drive to the muscles of 15 young and 15 elderly volunteers. The maximal rate of torque development as well as rate of torque development, impulse and neuromuscular activation in the early phase of contraction were determined. Spinal excitability was estimated using the H reflex technique. Changes at the muscle level were evaluated by analyzing the contractile properties and lean mass.The age-related decrease in iMVT was accompanied by a decline in voluntary activation and normalized surface EMG amplitude. Mechanical parameters of explosive voluntary strength were reduced while the corresponding muscle activation remained primarily unchanged. The spinal excitability of the vastus medialis was not different while M wave latency was longer. Contractile properties and lean mass were reduced.In conclusion, the age-related decline in iMVT of the quadriceps muscle might be due to a reduced neural drive and changes in skeletal muscle properties. The decrease in explosive voluntary strength seemed to be more affected by muscular than by neural changes.     

Neuromuscular control of scapula muscles during a voluntary task in subjects with Subacromial Impingement Syndrome. A case-control study

Imbalance of neuromuscular activity in the scapula stabilizers in subjects with Subacromial Impingement Syndrome (SIS) is described in restricted tasks and specific populations. Our aim was to compare the scapular muscle activity during a voluntary movement task in a general population with and without SIS (n = 16, No-SIS = 15).Surface electromyography was measured from Serratus anterior (SA) and Trapezius during bilateral arm elevation (no-load, 1 kg, 3 kg). Mean relative muscle activity was calculated for SA and the upper (UT) and lower part of trapezius (LWT), in addition to activation ratio and time to activity onset. In spite of a tendency to higher activity among SIS 0.10–0.30 between-group differences were not significant neither in ratio of muscle activation 0.80–0.98 nor time to activity onset 0.53–0.98.The hypothesized between-group differences in neuromuscular activity of Trapezius and Serratus was not confirmed. The tendency to a higher relative muscle activity in SIS could be due to a pain-related increase in co-activation or a decrease in maximal activation. The negative findings may display the variation in the specific muscle activation patterns depending on the criteria used to define the population of impingement patients, as well as the methodological procedure being used, and the shoulder movement investigated.     

The mechanism for the activation of latent TGF-beta during co-culture of endothelial cells and smooth muscle cells: cell-type specific targeting of latent TGF-beta to smooth muscle cells

Transforming growth factor-beta (TGF-beta) is secreted in a latent form and activated during co-culture of endothelial cells and smooth muscle cells. Plasmin located on the surface of endothelial cells is required for the activation of latent TGF-beta (LTGF-beta) during co-culture, and the targeting of LTGF-beta to the cellular surface is requisite for its activation. In the present study, the cellular targeting of LTGF- beta was examined. We detected the specific binding of 125I-large LTGF- beta 1 isolated from human platelets to smooth muscle cells but not to endothelial cells. A mAb against the latency-associated peptide (LAP) of large LTGF-beta 1 complex, which blocked the binding of 125I-large LTGF-beta 1 to smooth muscle cells, inhibited the activation of LTGF- beta during co-culture. The binding of 125I-large LTGF-beta 1 could not be competed either by mannose-6-phosphate (300 microM) or by the synthetic peptide Arg-Gly-Asp-Ser (300 micrograms/ml). These results indicate that the targeting of LTGF-beta to smooth muscle cells is required for the activation of LTGF-beta during co-culture of endothelial cells and smooth muscle cells. The targeting of LTGF-beta to smooth muscle cells is mediated by LAP, and the domain of LAP responsible for the targeting to smooth muscle cells may not be related to mannose-6-phosphate or an Arg-Gly-Asp sequence, both of which have been previously proposed as candidates for the cellular binding domains within LAP.     

The resilience of the size principle in the organization of motor unit properties in normal and reinnervated adult skeletal muscles

Henneman's size principle relates the input and output properties of motoneurons and their muscle fibers to size and is the basis for size-ordered activation or recruitment of motor units during movement. After nerve injury and surgical repair, the relationship between motoneuron size and the number and size of the muscle fibers that the motoneuron reinnervates is initially lost but returns with time, irrespective of whether the muscles are self- or cross-reinnervated by the regenerated axons. Although the return of the size relationships was initially attributed to the recovery of the cross-sectional area of the reinnervated muscle fibers and their force per fiber, direct enumeration of the innervation ratio and the number of muscle fibers per motoneuron demonstrated that a size-dependent branching of axons accounts for the size relationships in normal muscle, as suggested by Henneman and his colleagues. This same size-dependent branching accounts for the rematching of motoneuron size and muscle unit size in reinnervated muscles. Experiments were carried out to determine whether the daily amount of neuromuscular activation of motor units accounts for the size-dependent organization and reorganization of motor unit properties. The normal size-dependent matching of motoneurons and their muscle units with respect to the numbers of muscle fibers per motoneuron was unaltered by synchronous activation of all of the motor units with the same daily activity. Hence, the restored size relationships and rematching of motoneuron and muscle unit properties after nerve injuries and muscle reinnervation sustain the normal gradation of muscle force during movement by size-ordered recruitment of motor units and the process of rate coding of action potentials. Dynamic modulation of size of muscle fibers and their contractile speed and endurance by neuromuscular activity allows for neuromuscular adaptation in the context of the sustained organization of the neuromuscular system according to the size principle.     

Effects of two devices on the surface electromyography responses of eleven shoulder muscles during Azarian in gymnastics

Difficult elements of strength such as Azarian must be presented on the rings. Specific-muscles training may be realized with 2 devices, the Herdos and the Belt, both of which reproduce the competitive situation and allow many repetitions. The purpose of this study was therefore to compare the shoulder muscle activity during the performance of Azarian with each device. Our results show that muscles rhomboid, supraspinatus, deltoid (anterior, middle, and posterior parts), biceps brachii, and triceps brachii have significant (p < 0.05) higher root mean square value when gymnasts use the Belt compared with the Herdos. Although the Herdos and the Belt reproduce competitive movement, their muscle activities are quite different. The Herdos reduces the stress on the shoulder and elbow joints, whereas the Belt induces higher muscle activity and probably provides closer muscle synergisms to the rings.     

Movement of DNA sequence recognition domains between non-orthologous proteins

Comparisons of proteins show that they evolve through the movement of domains. However, in many cases, the underlying mechanisms remain unclear. Here, we observed the movements of DNA recognition domains between non-orthologous proteins within a prokaryote genome. Restriction–modification (RM) systems, consisting of a sequence-specific DNA methyltransferase and a restriction enzyme, contribute to maintenance/evolution of genomes/epigenomes. RM systems limit horizontal gene transfer but are themselves mobile. We compared Type III RM systems in Helicobacter pylori genomes and found that target recognition domain (TRD) sequences are mobile, moving between different orthologous groups that occupy unique chromosomal locations. Sequence comparisons suggested that a likely underlying mechanism is movement through homologous recombination of similar DNA sequences that encode amino acid sequence motifs that are conserved among Type III DNA methyltransferases. Consistent with this movement, incongruence was observed between the phylogenetic trees of TRD regions and other regions in proteins. Horizontal acquisition of diverse TRD sequences was suggested by detection of homologs in other Helicobacter species and distantly related bacterial species. One of these RM systems in H. pylori was inactivated by insertion of another RM system that likely transferred from an oral bacterium. TRD movement represents a novel route for diversification of DNA-interacting proteins.     

Neuromuscular efficiency of the rectus abdominis differs with gender and sport practice

The purpose of this investigation was to distinguish the abilities of the rectus abdominis (RA) muscle according to gender and sport training by means of neuromuscular parameters extracted from electromyography (EMG)-torque relationships. Thirty-eight healthy students, divided into 4 groups (i.e., 8 male runners, 10 female gymnasts, 12 male controls, and 8 female controls) were asked to perform 6 seconds of isometric trunk flexions at 20%, 25%, 75%, and 100% of their maximal voluntary contraction. Flexion torque and surface EMG of the RA muscle were recorded simultaneously to construct a EMG-torque relationship. Under maximal and submaximal conditions, an index of neuromuscular efficiency (NME) was determined to characterize the capacity of the RA muscle to develop a torque. At each level of contraction, the area of data scattering (ADS), reflecting torque and EMG fluctuations, was computed to express the capacity to maintain a constant target torque. Flexion torque, NME, and ADS values differed significantly between genders, but when data were related to anthropometric characteristics, no difference was observed. Although runners were not distinguished from male controls, gymnasts had higher flexion torque, higher NME, and lower ADS values than female controls had. These differences should reflect neural and muscular adaptations linked to the specificity of gymnastic training. These findings revealed different functional abilities of the RA muscle, according to gender and sport practices. The indices of neuromuscular capacities used in this study could constitute complementary tools to athletic trainers and professionals in sports medicine for evaluating and following, during sport-specific training programs, the abdominal muscle performance implied in force transfers with a lower cost and lower risks of back pain.     

Mechanical demand and multijoint control during landing depend on orientation of the body segments relative to the reaction force.

The purpose of this study was to determine how diverse momentum conditions and anatomical orientation at contact influences mechanical loading and multijoint control of the reaction force during landings. Male collegiate gymnasts (n=6) performed competition style landings (n=3) of drop jumps, front saltos, and back saltos from a platform (0.72 m) onto landing mats (0.12 m). Kinematics (200 fps), reaction forces (800 Hz) and muscle activation patterns (surface EMG, 1600 Hz) of seven lower extremity muscles were collected simultaneously. Between-task differences in segment orientation relative to the reaction force contributed to significant between-task differences in knee and hip net joint moments (NJM) during the impact phase. During the stabilization phase, ankle, knee, and hip NJMs acted to control joint flexion. Between-task differences in muscle activation patterns indicated that gymnasts scaled biarticular muscle activation to accommodate for between-task differences in NJM after contact. Activation of muscles on both sides of the joint suggests that impedance like control was used to stabilize the joints and satisfy the mechanical demand imposed on the lower extremity. Between-subject differences in the set of muscles used to control total body center of mass (TBCM) trajectory and achieve lower extremity NJMs suggests that control of multijoint movements involving impact needs to incorporate mechanical objectives at both the total body and local level. The functional consequences of such a control structure may prove to be an asset to gymnasts, particularly when required to perform a variety of landing tasks under a variety of environmental constraints.     

Ascomycota Members Dominate Fungal Communities during Straw Residue Decomposition in Arable Soil

This study investigated the development of fungal community composition in arable soil during the degradation of straw residue. We explored the short-term responses of the fungal community over 28 days of decomposition in soil using culture-independent polymerase chain reaction in combination with a clone library and denaturing gradient gel electrophoresis (DGGE). Fungal cellobiohydrolase I (cbhI) genes in the soil were also characterized, and their diversity suggested the existence of a different cellulose decomposer. The DGGE profiles based on fungal internal transcribed spacer analysis showed different successions of fungal populations during residue decomposition. Members of Lecythophora and Sordariales were dominant in the early succession, while Hypocrea and Engyodontium were better adapted in the late succession. The succession of fungal communities might be related to changes of residue quality during decomposition. Collectively, sequences assigned to Ascomycota members were dominant at different stages of the fungal succession during decomposition, revealing that they were key drivers responsible for residue degradation in the arable soil tested.     

What drives activation-dependent shifts in the force–length curve?

Skeletal muscles are rarely recruited maximally during movement. However, much of our understanding of muscle properties is based on studies using maximal activation. The effect of activation level on skeletal muscle properties remains poorly understood. Muscle optimum length increases with decreased activation; however, the mechanism responsible is unclear. Here, we attempted to determine whether length-dependent calcium effects, or the effect of absolute force underpin this shift. Fixed-end contractions were performed in frog plantaris muscles at a range of lengths using maximal tetanic (high force, high calcium), submaximal tetanic (low force, high calcium) and twitch (low force, low calcium) stimulation conditions. Peak force and optimum length were determined in each condition. Optimum length increased with decreasing peak force, irrespective of stimulation condition. Assuming calcium concentration varied as predicted, this suggests that absolute force, rather than calcium concentration, underpins the effect of activation level on optimum length. We suggest that the effect of absolute force is due to the varying effect of the internal mechanics of the muscle at different activation levels. These findings have implications for our understanding of in vivo muscle function and suggest that mechanical interactions within muscle may be important determinants of force at lower levels of activation.     

Electromyography variables during the golf swing: A literature review

The aim of the study was to review systematically the literature available on electromyographic (EMG) variables of the golf swing. From the 19 studies found, a high variety of EMG methodologies were reported. With respect to EMG intensity, the right erector spinae seems to be highly activated, especially during the acceleration phase, whereas the oblique abdominal muscles showed moderate to low levels of activation. The pectoralis major, subscapularis and latissimus dorsi muscles of both sides showed their peak activity during the acceleration phase. High muscle activity was found in the forearm muscles, especially in the wrist flexor muscles demonstrating activity levels above the maximal voluntary contraction. In the lower limb higher muscle activity of the trail side was found. There is no consensus on the influence of the golf club used on the neuromuscular patterns described. Furthermore, there is a lack of studies on average golf players, since most studies were executed on professional or low handicap golfers.Further EMG studies are needed, especially on lower limb muscles, to describe golf swing muscle activation patterns and to evaluate timing parameters to characterize neuromuscular patterns responsible for an efficient movement with lowest risk for injury.     

Mechanical energetic processes during the giant swing before the Tkatchev exercise

The goal of this study was to examine the possibility of the utilization of high bar and uneven parallel bar elasticity by the gymnasts through muscular work during the giant swing before the Tkatchev exercise on the high bar or uneven parallel bars. The performances were gathered during the Gymnastic World Championship in 1994. The set up consisted of two video cameras (50Hz) and two force measuring bars (500Hz). Twenty giant swings before the Tkatchev exercise, nine giant swings before the Tkatchev exercise after Tkatchev on the high bar and 15 giant swings before the Tkatchev exercise on the uneven parallel bars were analyzed. The giant swings were performed by 20 male and 15 female gymnasts. There are three phases during the giant swing exercise before the Tkatchev in which the systems (high bar-human body) total energy can be changed. During the first phase, energy is transferred from the gymnast's body into the bar. A clearly effective use of the bar's elasticity during the first phase could not be found. During the second phase, energy is transferred from the bar's back into the gymnast's body whose total energy increases. An increase in the energy of the system can only be achieved through muscular work. During the second phase of the various giant swing techniques no significant (p<0.05) difference in the energy increase through muscular work could be found. During the third phase, energy is once again produced by the gymnast through extension at the hip and shoulder joints.     

The interaction between mobile DNAs and their hosts in a fluctuating environment

The interaction between mobile DNA sequences and their hosts raises important questions in the context of hosts which reproduce clonally with only rare horizontal transmission between clones. The activity of some mobile DNAs as reversible mutators of genes raises the possibility that, in a fluctuating environment, cells may gain an advantage if they have mobile DNAs which mutate genes whose inactivation is favoured in one of the environments that the population encounters. Here we analyse a model of this process and ask what would be the optimal rate of transposition in a population whose elements are maintained by this mechanism. We also examine the impact of horizontal transfer on such a population. With movement of elements between cells, we can imagine elements with differing rates of transposition and host cells with differing rates of transposition. We find that evolution in the population of elements favours a rapid rate of transposition, and evolution of the host cells favours cells in which this rapid rate of element-dependent transposition results in an optimal rate of transposition per cell. However, when horizontal transfer rates are high, some unexpected features of the model are observed. In particular, a polymorphism between cell types (some with an optimal rate of transposition and some with no transposition at all from endogenous elements) can be stably maintained. We consider the possible biological predictions of this analysis.     

Mobile DNA can drive lineage extinction in prokaryotic populations

Natural selection ultimately acts on genes and other DNA sequences. Adaptations that are good for the gene can have adverse effects at higher levels of organization, including the individual or the population. Mobile genetic elements illustrate this principle well, because they can self‐replicate within a genome at a cost to their host. As they are costly and can be transmitted horizontally, mobile elements can be seen as genomic parasites. It has been suggested that mobile elements may cause the extinction of their host populations. In organisms with very large populations, such as most bacteria, individual selection is highly effective in purging genomes of deleterious elements, suggesting that extinction is unlikely. Here we investigate the conditions under which mobile DNA can drive bacterial lineages to extinction. We use a range of epidemiological and ecological models to show that harmful mobile DNA can invade, and drive populations to extinction, provided their transmission rate is high and that mobile element‐induced mortality is not too high. Population extinction becomes more likely when there are more elements in the population. Even if elements are costly, extinction can still occur because of the combined effect of horizontal gene transfer, a mortality induced by mobile elements. Our study highlights the potential of mobile DNA to be selected at the population level, as well as at the individual level.     

Crystal Structure of the Leucine Aminopeptidase from Pseudomonas putida Reveals the Molecular Basis for its Enantioselectivity and Broad Substrate Specificity

The zinc-dependent leucine aminopeptidase from Pseudomonas putida (ppLAP) is an important enzyme for the industrial production of enantiomerically pure amino acids. To provide a better understanding of its structure-function relationships, the enzyme was studied by X-ray crystallography. Crystal structures of native ppLAP at pH 9.5 and pH 5.2, and in complex with the inhibitor bestatin, show that the overall folding and hexameric organization of ppLAP are very similar to those of the closely related di-zinc leucine aminopeptidases (LAPs) from bovine lens and Escherichia coli. At pH 9.5, the active site contains two metal ions, one identified as Mn²⁺ or Zn²⁺ (site 1), and the other as Zn²⁺ (site 2). By using a metal-dependent activity assay it was shown that site 1 in heterologously expressed ppLAP is occupied mainly by Mn²⁺. Moreover, it was shown that Mn²⁺ has a significant activation effect when bound to site 1 of ppLAP. At pH 5.2, the active site of ppLAP is highly disordered and the two metal ions are absent, most probably due to full protonation of one of the metal-interacting residues, Lys267, explaining why ppLAP is inactive at low pH. A structural comparison of the ppLAP-bestatin complex with inhibitor-bound complexes of bovine lens LAP, along with substrate modelling, gave clear and new insights into its substrate specificity and high level of enantioselectivity.