Ultrastructure of “Gordona aurantiaca” Tsukamura 1971

Ultrastructure of Gordona aurantiaca^* M 296 (8128) was studied after the lead citrate coloration, whereas the cell envelope architecture was investigated by ruthenium red staining for outer wall acidic polysaccharides and the periodic-acid-thiocarbohydrazide-silver-proteinate cytochemical procedure (Thiéry method) for the detection of 1-2 glycol bond containing polysaccharides. The ultrastructural morphology of bacteria was distinct from both the mycobacteria and nocardia. The bacilli had a typical gram-positive cell wall that contained a thin, uniformly distributed, polysaccharide outer layer (POL) at its surface. The Thiéry cytochemical method stained only the cytoplasmic membrane, but not the cell wall, a feature that is common to the mycolic acid containing theCorynebacterium-Mycobacterium-Nocardia (CMN) group of organisms. The negative staining of the unfixed preparations of bacilli showed ribbonlike surface structures, common to the CMN group of organisms. The electron-microscopic preparations showed numerous lysing bacilli with bacteriophages indicating that the strain used was lysogenic.     

A framework for classifying subsistence production types of Nepal

This paper defines a four-tiered, hierarchical system for classifying subsistence production types in Nepal and uses the system to describe the subsistence communities of central Nepal. The system first divides Nepal into a grid of homogenous parts or cells, second, it divides, the cells into ridge-valley slope sequences, third, defines production types for the ridge-valley slope sequences, and, finally, distinguishes local variants of the production types. After describing these tiers, I apply these categories to the northern portions of the central and western development regions where I have done research. I then define how representatives of Nepal as a whole are the production types defined for the central and western development regions. Finally, the paper locates some completed studies within its framework.     

Hypoxia Tolerance of Two Haplochromine Cichlids: Swamp Leakage and Potential for Interlacustrine Dispersal

The ability to tolerate hypoxia in some haplochromine cichlid fishes contributes to the richness of habitats occupied by the lineage and may be important in interlacustrine dispersal through swampy channels. Lacustrine members of the genus Astatotilapia tend to be ecologically plastic but are rarely encountered in the interior of dense swamps. A notable exception is seen in the swamp corridor that joins Lake Kabaleka with Lake George, Uganda, where one species (Astatotilapia wrought-iron) is abundant, and a second species, A. aeneocolor, is rare. Both species are abundant in the open waters of the main lake. In this paper, we compare physiological (oxygen consumption) and behavioral indicators of hypoxia tolerance between A. wrought-iron from swamp and open-water habitats and between the two species of Astatotilapia. When exposed to progressive hypoxia, all fish used aquatic surface respiration (ASR); however, swamp-dwelling A. wrought-iron showed lower gill ventilation rates prior to the initiation of ASR, higher pre-ASR aggression rates, higher swimming speed during ASR, and a higher rate of bubble exchange than both the open-water group of A. wrought-iron and A. aeneocolor. These differences may reflect interpopulational variation in selection pressure for low-oxygen tolerance between swamp and open-water habitats. Several lines of evidence suggest that A. wrought-iron was in general more hypoxia tolerant than A. aeneocolor. These include a lower ASR₉₀ threshold, a drop in gill ventilation rate with the onset of ASR, and lower rate of equilibrium loss under extreme hypoxia in A. wrought-iron. The routine metabolic rate and critical oxygen tension did not differ between swamp-dwelling and open-water A. wrought-iron, or between A. wrought-iron and A. aeneocolor. Comparative data on the ASR thresholds and critical oxygen tensions of the Astatotilapia species from Lake Kabaleka and other East African cichlids suggest intermediate hypoxia tolerance. Nevertheless, our study suggests that some generalized lacustrine haplochromines may leak through swamp corridors even under relatively extreme conditions.     

The ultrastructure of holdfasts, “rhizoids”, and “slime tracks” in thraustochytriaceous fungi and Labyrinthula spp.

Summary The fine structure of the holdfasts or rhizoids is described for the thraustochytriaceous organisms, Thraustochytrium motivum, Schizochytrium aggregatum, and an unidentified organism, denoted T-20, which resembles S. aggregatum and Labyrinthula spp. Labyrinthula algeriensis and L. minuta slime track ultrastructure is also described. The holdfasts, rhizoids, and tracks have the same basic fine structure and are collectively termed ectoplasmic nets. They are delimited by a unit membrane which is in continuity with the plasmalemma, contain no cytoplasmic organelles only membrane-limited cisternae, and contain a fibrogranular ground substance. The nets appear to arise from one or as many as 20 organelle complexes consist of an approximately disk-shaped electron-dense granular aggregate in which are embedded portions of cisternae of the endoplasmic reticulum or perinuclear clear continuum. The cisternae appear to contribute small (ca. 17 nm diameter) vesicles to the granular aggregate which coalesce to form internal membranes of the net elements. The sagenogenetosome underlies the plasmalemma where it evaginates to form the delimiting membrane of the main trunk element of the net. No continuous membrane separates the net contents from the cytoplasm, only the granular aggregate.In L. algeriensis, L. minuta, and T-20 the net is necessary for motility of nonflagellated, nonamoeboid cells. Presence of the nets is not associated with motility in S. aggregatum and T. motivum. The possible taxonomic significance of the observations is discussed.Contribution No. 456, Virginia Institute of Marine Science, Gloucester Point, Virginia 23062.Supported in part by the Oceanography Section, National Science Foundation, NSF Grant GA-31014.     

Macromolecular differentiation of Golgi stacks in root tips ofArabidopsis andNicotiana seedlings as visualized in high pressure frozen and freeze-substituted samples

Summary The plant root tip represents a fascinating model system for studying changes in Golgi stack architecture associated with the developmental progression of meristematic cells to gravity sensing columella cells, and finally to young and old, polysaccharideslime secreting peripheral cells. To this end we have used high pressure freezing in conjunction with freeze-substitution techniques to follow developmental changes in the macromolecular organization of Golgi stacks in root tips ofArabidopsis andNicotiana. Due to the much improved structural preservation of all cells under investigation, our electron micrographs reveal both several novel structural features common to all Golgi stacks, as well as characteristic differences in morphology between Golgi stacks of different cell types.Common to all Golgi stacks are clear and discrete differences in staining patterns and width of cis, medial and trans cisternae. Cis cisternae have the widest lumina (30 nm) and are the least stained. Medial cisternae are narrower (20 nm) and filled with more darkly staining products. Most trans cisternae possess a completely collapsed lumen in their central domain, giving rise to a 4–6 nm wide dark line in cross-sectional views. Numerous vesicles associated with the cisternal margins carry a non-clathrin type of coat. A trans Golgi network with clathrin coated vesicles is associated with all Golgi stacks except those of old peripheral cells. It is easily distinguished from trans cisternae by its blebbing morphology and staining pattern. The zone of ribosome exclusion includes both the Golgi stack and the trans Golgi network.Intercisternal elements are located exclusively between trans cisternae of columella and peripheral cells, but not meristematic cells. In older peripheral cells only trans cisternae exhibit slime-related staining. Golgi stacks possessing intercisternal elements also contain parallel rows of freeze-fracture particles in their trans cisternal membranes. We propose that intercisternal elements serve as anchors of enzyme complexes involved in the synthesis of polysaccharide slime molecules to prevent the complexes from being dragged into the forming secretory vesicles by the very large slime molecules. In addition, we draw attention to the similarities in composition and apparent site of synthesis of xyloglucans and slime molecules.Dedicated to the memory of Professor Oswald Kiermayer     

The polypeptide structure and assembly of Ly-2/3 heterodimers

Mild reduction of mature, thymic Ly-2/3 heterodimers of M _r 67 000 resulted in dissociation into three individual polypeptide chains, , , and , of respective M _r values 38000, 35000, and 30000. The and chains were both immunoprecipitated by a monoclonal antibody directed to the Ly-2.1 epitope whereas the Ly-3.1 antibody bound only the chain. The possibility that the and chains of each heterodimer established their interchain links within a labile precursor protein in which a and segments were fused was considered but discounted by the finding that in mice heterozygous for both Ly-2 and Ly-3 loci, the Ly-2 product of one chromosome was not exclusively joined to Ly-3 structures coded by the same chromosome. By utilizing ionic detergents which selectively alter the charge of intrinsic membrane proteins, both Ly-2 and Ly-3 polypeptides were shown to have membrane insertion sites. It is suggested that as a consequence of their likely synthesis on membrane-bound polysomes, newly synthesized Ly-2 and Ly-3 structures accumulate within the same subcellular compartment — the membranes of the rough endoplasmic reticulum. Their elevated concentration within this space may facilitate a low affinity binding interaction between Ly-2 and Ly-3 which is later stabilized by interchain disulfide bond formation.Abbreviations used in this paper BSA bovine serum albumin - DOC sodium deoxycholate - DTT dithiothreitol - HA hemagglutinin - HTAB hexadecyltrimethylammonium bromide - RER rough endoplasmic reticulum - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - TX100 Triton X-100     

SDS Induced Structural Changes in α-Crystallin and It’s Effect on Refolding

-Crystallin, a major eye lens protein and a key member of the small heat shock protein family, acts like a chaperone by preventing aggregation of substrate proteins. One of the hallmarks of most small heat shock proteins is their existence as a large oligomer, the role of which in its function is not understood at present. We have studied the role of the oligomer in the stability of its structure against SDS induced destabilization by CD measurements. -Crystallin from bovine source as well as recombinant preparation was used for this purpose. As SDS concentration was gradually increased, the -sheet structure was diminished followed by concomitant increase in the -helical structure. The quaternary structural changes in presence of SDS were also monitored by light scattering, polarization and anisotropy measurements. It was found that the breakdown of the oligomeric structure was nearly complete above 1 mM SDS concentration. The results were compared with that of a monomeric -crystallin, which is also a major -sheet protein like -crystallin. When -crystallin was first converted into monomeric random coil structure in presence of 6 M urea and allowed to refold in SDS solution, amount of -helix was more than that incubated directly in the same concentration of SDS. The results show that -crystallin attains extra structural stability against external stress due to its oligomeric structure. The implication for the extra stability is discussed in reference to its function as molecular chaperone.     

Ultrastructural analysis of the sperm cells of mature pollen of maize,Zea mays

Summary Ultrastructural analysis of the mature viable unhydrated pollen of maize,Zea mays from dehiscent anthers shows that the sperm cells are physically distant, each bounded by an envelope comprising their own plasma membrane and the inner plasma membrane of the vegetative cell. The chondriome is unusual in containing one or more filamentous complexes, up to 12m in length appressed to the side of the sperm nucleus. The extensions at each end of the elongate sperm cells contain longitudinally-oriented arrays of endoplasmic lamellae. In a three-dimensional reconstruction of serial thin sections, there is a long J-shaped sperm, c. 35 × 5m and up to 1m in thickness, sited within pointed evaginations of the vegetative nucleus and a second shorter sperm c. 20 × 5m and up to 3.5m in thickness.Abbreviations PA-TCH-SP periodic acid-thiocarbohydrazide-silver proteinate - DAPI 4,6-diamino-2-phenylindole - SC sperm cell - Sn sperm nucleus - Ua-Pb Uranyl acetate-lead citrate staining - ER endoplasmic reticulum     

A phylogenetic analysis ofLeucaena (Leguminosae: Mimosoideae)

Chloroplast DNA restriction fragment length polymorphisms have been used to reconstruct the maternal phylogeny of all the known taxa in the small neotropical legume genusLeucaena. Three major plastome clades were recognized, but these did not conform with relationships between the taxa proposed on other characters from morphology, cytology or hybridization. The maternal parentage of tetraploids within the genus has been proposed. Evidence for introgression was found between diploidL. diversifolia and tetraploidL. diversifolia. The implications of these results for the origin of the cultivated taxa are discussed.     

Supernatants derived from CD8+ lymphocytes activated by mycobacterial fractions inhibit cytokine production. The role of interleukin-6

Our study examined the effects of supernatants derived from CD8+ lymphocytes treated with high molecular weight components ofMycobacterium tuberculosis on cytokine production. Such suppressor but not control supernatants increased the production of IL-4 and IL-6 whilst suppressing IL-1, TNF-alpha, IL-2 and IFN- productionby monocytes andlymphocytes. The effects on cytokine production were time dependent being observed as early as 4 hours with peak activity observed at 24 hours.The inhibition of IL-1 and TNF-alpha by monocytes appeared to be related to increases in IL-6 levels present in supernatants of non-adherent lymphocytes incubated with mycobacterial components. This was confirmed by studies demonstrating that the addition of recombinant IL-6 to cultures depressed the production of these cytokines. Furthermore the addition of monoclonal anti-IL6 to such cultures restored the production of IL-1 and TNF-alpha. The results suggest that mycobacterial components inhibit host cellular functions by manipulating the host's cytokine network.     

Effect of 17β-estradiol on cells stained for FSHβ and/or LHβ in the dog pituitary gland

Summary The effect of long-term treatment (52 weeks) with high doses of 17-estradiol (1.28 mg/kg/week intramuscularly) on gonadotrophs was studied in the pituitary gland of the beagle bitch. For immunochemical staining the immunoperoxidase technique and antisera to the specific beta () subunits of FSH and LH were employed. For control purposes antisera to the following hormones were also used: bovine TSH, canine GH, canine PRL and porcine ACTH¹. In the pars distalis and pars tuberalis of control bitches, in addition to the cells which react solely with antisera to either LH or FSH, most cells were reactive to both antisera. The cells stained for FSH were less numerous than those shown to contain LH. TSH, PRL, GH and ACTH/MSH were localized in distinctly different cell types in the pars distalis of all control animals. In the treated bitches, almost complete regression of cells classically identified as gonadotrophs and stained for LH was observed. On the other hand, using the antiserum to FSH, selective immunochemical staining was localized in cells fitting the morphological characteristics of TSH cells. All these cells were also stained for TSH. However, a few cells were also shown to react solely with the antiserum to TSH. These cells, which seem to contain both TSH and FSH, were further clearly differentiated from PRL, GH and ACTH/MSH cells on the basis of their cytological features, intraglandular distribution and by immunochemical double staining. These observations support the concept that the one cell-one hormone theory may not necessarily apply to the glycoprotein hormones of the dog pituitary gland.Abbreviations of Hormones cited in this Paper ACTH Adrenocorticotropin - FSH Follicle Stimulating Hormone - GH Growth Hormone - LH Luteinizing Hormone - MSH Melanocyte Stimulating Hormone - PRL Prolactin - TSH Thyrotropin The authors are grateful to Mrs. K. Oertel for carrying out the experimental work on animals, to Mrs. B. Schilk and Miss U. Tüshaus for their excellent technical assistance, and to Dr. P. Günzel for his advice and encouragement     

Mode of uptake of sterol by Arthrobacter simplex

The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M _r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10^–3g l^–1h^–1) could not adequately account for the rate of sterol uptake (72×10^–3g l^–1h^–1) and the specific growth rate (56×10^–3 h^–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm^–1), the sterol-grown cells had strong surface activity (40mNm^–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.     

A pressure test method monitors biofilm coated particle fluidization

A fluidized bed denitrifying reactor was run to examine the vertical segregation of sand particles on the basis of different biofilm coverage, so far neglected when modelling fluidized beds. The segregation was found to be significant and it can be directly correlated with the vertical hydrostatic pressure profile in the bed. A procedure was developed for the rapid determination of biofilm thickness from hydrostatic pressure data using a recently published method based on the use of the novel criteria expansion coefficient and specific occupied volume. A key feature of the procedure is the particle content, which can be calculated from particle characteristics and is correlated in this study with the hydrostatic pressure gradient. The method was verified by directly measuring biofilm thickness as a function of the vertical position in the bed. This way biofilm thickness can be calculated from a readily measurable hydrostatic pressure profile with an error of 0.04–0.06 mm. This error is believed to be due to N₂ gas entrapment in the denitrifying biofilm and to the original inaccuracy of the determination of particle size and volume. The method is rather insensitive to the exact biofilm density when the usual high-density carrier material is used.     

The β-1,3-glucan-binding protein from the crayfish Pacifastacus leniusculus,when reacted with a β-1,3-glucan,induces spreading and degranulation of crayfish granular cells

Summary A -1,3-glucan-binding protein (GBP) was purified from crayfish plasma, and incubated with laminarin (L), a -1,3-glucan. The GBP reacted with laminarin (GBP-L) induced strong spreading and partial degranulation of isolated and separated crayfish granular haemocytes. However, neither the GBP nor laminarin alone induced any changes in the crayfish granular cells. When monolayers of granular haemocytes were incubated with 20 g of GBP-L, more than 82% of the haemocytes were affected. The activity of GBP-L on granular cells was dose-dependent and a plateau was reached at 10 g of GBP-L. The degranulation of crayfish haemocytes induced by GBP-L seemed to occur by a regulated exocytosis, since it was strongly inhibited by specific blockers of this process such as SITS or calmidazolium. Monospecific anti-GBP antibodies also totally blocked the effect of GBP-L on crayfish granular cells. Indirect immunofluoresence staining demonstrated that the GBP-L could bind to the surface of granular cells, whereas GBP did not bind or bound very weakly to the haemocyte surface.     

Growth of a thermotolerant ethanol-producing strain of Kluyveromyces marxianus on cellobiose containing media

Summary The thermotolerant yeast strain, Kluyveromyces marxianus 1MB 3, was shown to be capable of limited growth on cellobiose containing media at 45°C. Growth, sugar utilization and ethanol production were shown to increase in the presence of exogenously added thermostable fungal -glucosidase. During active growth of the organism on cellobiose-containing media, -glucosidase activity was detected in cell lysate preparations with only minor amounts of activity present in the extracellular culture filtrate. The results suggest that limitations in ethanol production by this organism during growth on cellobiose containing media may be overcome by addition of exogenously added -glucosidase which results in increased substrate access to the biocatalytic unit.     

Average phase difference theory and 1∶1 phase entrainment in interlimb coordination

The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency _c was varied by a metronome, and scaled to the eigenfrequency _v of the coupled system K was assumed to vary inversely with _c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is _c/_v rather than _c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.     

Physiology and biochemistry of the stigmatic fluid of Petunia hybrida

Summary Production of stigma exudate per flower of Petunia hybrida is about 200 g. The effect of light, temperature, metabolic poison and emasculation on the production of the exudate at different ages of the bud has been studied. The presence of a thin film of water below the stigmatic exudate has been demonstrated. Physical properties of the exudate such as relative viscosity and surface tension have also been determined. Chemical analysis of the stigmatic fluid showed that it consists primarily of an oil, sugars and amino acids. No protein could be detected. It also contains no acid phosphatase.Behaviour of the pollen from its deposition on the stigmatic fluid until it germinates on the stigma surface has been studied in vivo and also with the aid of an artificial stigma.The role of the stigmatic fluid in pollination has been determined.

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Zusammenfassung Die Narbenflüssigkeit, welche von zahlreichen Pflanzen ausgeschieden wird zu einem Zeitpunkt, da die Blüte reif ist zur Bestäubung, spielt eine wichtige Rolle bei der Fixierung des Pollens. Um so überraschender ist die Tatsache, daß hinsichtlich der Physiologie und Biochemie des Narbenschleimes so wenig detaillierte Informationen zur Verfügung stehen.Bei Petunia wird je Blüte im Mittel 200 g Narben-Flüssigkeit produziert. Die Intensität der Narbenschleimproduktion ist abhängig von der Temperatur und der Länge der Lichtperiode. Hingegen wird die Ausscheidung der Narbenflüssigkeit durch Hemmung der Atmung und durch partielle Kastration nicht beeinflußt.Die physikalischen Eigenschaften (Dichte, relative Viscosität und Oberflächenspannung) werden bestimmt. Bei der chemischen Analyse des Narbenschleimes ergab sich, daß dieser hauptsächlich aus einem öligen Fett besteht, dessen Fettsäure-Zusammensetzung ermittelt wurde. Hingegen werden nur sehr geringe Spuren von Zuckern und Aminosäuren gefunden. Der Narbenschleim ist frei von Proteinen und zeigt keine enzymatische Aktivität.Das Verhalten des Pollens bei der Landung in einem Tropfen der Narbenflüssigkeit wird in vivo und mit Hilfe einer künstlichen Modell-Narbe untersucht.Die Bedeutung der Narbenflüssigkeit im Zusammenhang mit der Bestäubung und den einleitenden Stadien der Pollenkeimung wird diskutiert.

     

Characterization of the fibrillar layer at the epithelial-mesenchymal junction in tooth germs

A characteristic layer containing numerous fibrils is associated with the basement membrane of the inner enamel epithelium during the early stages of odontogenesis. However, its nature is not well understood. In this study, the layer was examined with high-resolution electron microscopy and immuno-histochemical staining. Tooth germs of monkeys (Macaca fuscata) were studied and each fibril in the layer was found to be a tubular structure, 8–9 nm in width, resembling a basotubule, the tubular structure previously observed in various basement membranes. The space between the fibrils was filled with a network formed by irregular anastomosing strands with an average thickness of 4 nm; these strands resembled the cords forming the network in the lamina densa of basement membranes. After immunoperoxidase staining, fine threads immunoreactive for laminin staining were seen winding along the strands of the network, and 1.5-nm wide filaments, immunoreactive for type IV collagen, took the form of a network arrangement. The 5-nm-wide ribbon-like structures associated with the strands were identified as heparan sulfate proteoglycan by immunostaining. These results are similar to those obtained for the cord network of the lamina densa. The fibrillar layer therefore represents a highly specialized lamina fibroreticularis of the basement membrane of the inner enamel epithelium, and rich in basotubules.     

The cytology of phytophthora infestans

Summary Phytophthora infestans has three kinds of somatic nuclei: an oval shaped nucleus (approx. 3.1×2.7 ) which stains diffusely except for a crescent shaped Feulgen positive cap which stains intensely; a granular nucleus whose contents are organized into a fairly constant number of stained bodies, and, a deeply staining condensed nucleus. The capped nucleus is thought to be metabolic or resting and the granular nucleus is thought to be dividing as it is most commonly found in hyphal tips. Attenuated forms of all three kinds of nuclei are found.Nuclear division is mitotic and intranuclear. Eight—ten chromosomes are seen at metaphase.Sporangia have a mean of 6.3 nuclei which is constant for age and strain of culture. Sporangia become multinucleate as a result of nuclear migration and not by division in the developing sporangium. Zoospores are usually uninucleate.The nuclear cap is persistent throughout nuclear division when it also divides. It is associated with flagella production and nuclear migration and has some of the properties of a blepharoplast.     

Expression,purification, characterization,and deletion mutations of phosphorylase kinase γ subunit: identification of an inhibitory domain in the γ subunit

A catalytic fragment, _1-298, derived from limited chymotryptic digestion of phosphorylaseb kinase (Harris, W.R.et al., J. Biol. Chem., 265: 11740–11745, 1990), is reported to have about six-fold greater specific activity than does the subunit-calmodulin complex. To test whether there is an inhibitory domain located outside the catalytic core of the subunit, full-length wild-type and seven truncated forms of were expressed inE. coli. Recombinant proteins accumulate in the inclusion bodies and can be isolated, solubilized, renatured, and purified further by ammonium sulfate precipitation and Q-Sepharose column. Four out of seven truncated mutants show similar ( _1-353 and _1-341) or less ( _1-331 and _1-276) specific activity than does the full-length wild-type , _1-386. Three truncated forms, _1-316, _1-300, and _1-290 have molar specific activities approximately twice as great as those of the full-length wild-type and the nonactivated holoenzyme. All recombinant s exhibit similarK _m values for both substrates, i.e., about 18M for phosphorylaseb and about 75 M for MgATP. Three truncated s, _1-316, _1-300, and _1-290, have a 1.9- to 2.5-fold greater catalytic efficiency (V _max/K _m) than that of the full-length wild-type and a 3.5- to 4.5-fold greater efficiency than that of the truncated _1-331. This evidence suggests that there is at least one inhibitory domain in the C-terminal region of , which is located at _301-331· _1-290, but not _1-276, which contains the highly conserved kinase domain, is the minimum sequence required for the subunit to exhibit phosphotransferase activity. Both _1-290 and _1-300 have several properties similar to full-length wild-type , including metal ion responses (activation by free Mg²⁺ and inhibition by free Mn²⁺) pH dependency, and substrate specificities.