Calmodulin and the target size of the (Ca2+ + Mg2+)-ATPase of human red-cell ghosts

An average target size of 251 kDa has been obtained for the (Ca²⁺ + Mg²⁺)-ATPase of calmodulin-depleted erythrocyte ghosts by radiation inactivation with 16 MeV electrons. This is close to twice the size of the purified calcium-pump polypeptide. When calmodulin was included during the ATPase assay, a component of about 1 MDa appeared in addition to the activated dimer.     

Phase transition kinetics of phosphatidic acid bilayers A stopped-flow study of the electrostatically induced transition

The kinetics of the electrostatically induced phase transition of dimyristoyl phosphatidic acid bilayers was followed using the stopped-flow technique. The phase transition was triggered by a fast change in the pH or the magnesium ion concentration and followed by recording the time dependence of the absorbance. When the phase transition was induced by a pH jump the time course of the absorbance could be described by two exponentials, their time constants displaying the for cooperative processes characteristic maximum at the transition midpoint. The time constants are in the 10 and 100 ms range for the H⁺ triggered transition from the fluid to the ordered state. A third slower process shows no appreciable temperature dependence and is probably caused by vesicle aggregation. For the OH^--induced transition fron the ordered to the fluid state the time constants are in the 100 and 1000 ms range. The fluid-ordered transition could also be triggered by addition of magnesium ions. Of the several observed processes only the fastest in the 10–100 ms time range could definitely be assigned to the fluid-ordered transition while the others are due to aggregation phenomena. The experimental data were compared with results obtained from pressure jump experiments and could be interpreted on the basis of theories for non-equilibrium relaxation.     

Indol-3yl-acetic acid in roots of Zea mays

Indol-3yl-acetic acid was identified in extracts of sterile roots of Zeamays seedlings by means of TLC, chromogenic reactions, GLC and GC-MS.     

Membrane lipid composition and susceptibility to bile salt damage

Erythrocyte membranes with low sphingomyelin: choline-containing phospholipid ratios haemolyse at low concentrations of the bile salt, glycocholate. Erythrocytes with higher sphingomyelin: choline-containing phospholipid ratios require progessively greater concentrations of the bile salt for lysis.Sublytic concentrations of glycocholate remove phospholipid and acetylcholinesterase from the membranes. Membranes with low sphingomyelin: choline-containing phospholipid ratios lose both particulate (microvesicles of distinct composition) and ‘solubilized’ material, the particulate form predominating. The proportion of particulate material falls with increase of the membrane sphingomyelin: choline-containing phospholipid ratio and those membranes of highest sphingomyelin: choline-containing phospholipid ratio lose material predominantly in ‘solubilized’ form.Sheep erythrocytes treated to increase their content of phosphatidylcholine (and thereby reduce their membrane sphingomyelin: choline-containing phospholipid ratio) become more susceptible to lysis by glycocholate.These observations indicate a correlation between membrane lipid composition and the perturbation of membranes with bile salt; they also point to possible features of membranes capable of surviving exposure to the high bile salt concentrations of the biliary tract.     

The physiological basis of seed dormancy in Avena fatua. VIII. Action of malonic acid

Adkins, S. W., Symons, S. J. and Simpson, G. M. 1988. The physiological basis of seed dormancy in Avena fatua . VIII. Action of malonic acid - Physiol. Plant, 72: 477–482.
A low concentration of malonic acid (50 m M ) induced germination in four genetically pure dormant lines of Avena fatua L. Sensitivity to this treatment was poor immediately after harvest but increased markedly during after-ripening, indicating that the mode of action of malonic acid (50 m M ) was similar to that of another organic acid, citric acid. Over the concentration range (10–50 m M ) where malonic acid promoted germination, oxygen uptake was also stimulated, and this was before the first visible signs of germination. At higher concentrations (100–300 m M ) where there was no promotion of germination, malonic acid strongly inhibited oxygen uptake. These results show that malonic acid has a dual effect on oxygen uptake and subsequent germination. Low concentrations (10–50 m M ) act by stimulating the Krebs cycle and germination through an acidification reaction like citric acid, and high concentrations (100–300 m M ) act by inhibiting germination through enzymatic restraint of the Krebs cycle.
The stimulation of both oxygen uptake and germination by three established germination promoters (sodium nitrate, citric acid and ethanol) was inhibited by a high concentration of malonic acid (200 m M ) but unaffected by a low concentration (50 m M ). These results show that oxygen uptake, and hence the activity of the Krebs cycle, are important processes involved in the dormancy breaking mechanism of these three promotors.     

Marketing approval of mogamulizumab: A triumph for glyco-engineering

Therapeutic properties of antibodies strongly depend on the composition of their glycans. Most of the currently approved antibodies are produced in mammalian cell lines, which yield mixtures of different glycoforms that are close to those of humans, but not fully identical. Glyco-engineering is being developed as a method to control the composition of carbohydrates and to enhance the pharmacological properties of mAbs. The recent approval in Japan of mogamulizumab (POTELIGEO^®), the first glyco-engineered antibody to reach the market, is a landmark in the field of therapeutic antibodies. Mogamulizumab is a humanized mAb derived from Kyowa Hakko Kirin’s POTELLIGENT^® technology, which produces antibodies with enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) activity. The approval was granted April 30, 2012 by the Japanese Ministry of Health, Labour and Welfare for patients with relapsed or refractory CCR4-positive adult T-cell leukemia-lymphoma.     

Inhibition of 3,4-dihydroxy-l-phenylalanine decarboxylase in rat striatal synaptosomes by amino acids interacting with substrate transport

Dopamine synthesis from 3,4-dihydroxy-l-phenylalanine in rat striatal synaptosomes was inhibited by a number of amino acids with aromatic or large aliphatic side chains. Inhibition was not seen when aromatic amino acid decarboxylase activity was measured in disrupted synaptosomes. Similarly, inhibition of dopamine synthesis from tyrosine was seen in the presence of leucine. The inhibition most likely results from interactions of the amino acids with substrate transport across the synaptosome plasma membrane, rather than directly with the catalytic enzymes. The kinetic data obtained are used to infer information about the relevant transport process; they suggest the potential importance of amino acid efflux as a regulatory step.     

Sequential polypeptides: 5. Statistical mechanical analysis of helix-coil transition in sequential polypeptides

A statistical mechanical theory of the helix-coil transition in sequential polypeptides is developed assuming that the statistical weights of the Zimm-Bragg parameters of a given residue depend on the type of adjacent residues. In the case of a sequential polypeptide consisting of two kinds of residues, the theory describes the helix- coil transition of the polypeptide in terms of the Zimm-Bragg parameters associated with the corresponding residues. The theory is then used to determine this parameter, as a function of temperature, from experimental data for transition temperature as a function of solvent composition, for a series of sequential polypeptides consisting of Glu(OBzl) and Lys(Chz) residues in mixtures of dichloroacetic acid and 1,2-dichlorethane. This parameter is then combined with the Zimm-Bragg parameters for the parent homopolypeptides, and the theory used to predict helix coil transition curves which are in good agreement with the experimental ones for the sequential polypeptides studied.     

Ornithine transaminase from Cucurbita maxima cotyledons

During germination a marked increase in both soluble and particulate ornthine transaminase occurs in pumpkin cotyledons. Both enzymes had a pH optimum of 8.3 and a requirement for ornthine and α-ketoglutarate. Other keto acids or amino donors showed little activity. The enzymes required an active sulphydryl group for maximum activity. Exogenous pyridoxal phosphate was not required, but hydroxylamine inhibited the reaction and added pyridoxal phosphate overcame this inhibition. Proline inhibited the reaction and may play a role in the fate of ornithine in pumpkin cotyledons.     

Driving forces and current-voltage characteristics of amino acid transport in Riccia fluitans

The complete steady-state I–V relationship of α-aminoisobutyric acid transport across the plasmalemma of rhizoid cells from Riccia fluitans has been measured and analysed with special emphasis on α-aminoisobutyric acid equilibrium and saturation conditions. (A) The electrical data show that: (1) the amino acid-induced electrical current saturates after the addition of the amino acid, regardless of the concentration; (2) a steady state is reached 1–2 h after incubation in α-aminoisobutyric acid, but after less that 5 min in the presence of 1 mM CN⁻; (3) the steady-state I–V characteristic of α-aminoisobutyric acid transport is a sigmoid curve and fairly symmetric in current with respect to the voltage axis; and (4) the equilibrium potential is clearly a function of the amino acid accumulation ratio. It is suggested that the sigmoid curve represents the characteristic of carrier-mediated α-aminoisobutyric acid transport with a voltage-insensitive step, possibly the translocation of the unloaded carrier, rate-limiting. Since under normal conditions the voltage-sensitive rate constant k_oi is much greater than k_io, it is further suggested that the energy to drive this system is put into the transfer of positive charge from outside to the cytoplasm. (B) Accumulation ratios have been determined by inspection of current-voltage data, and additionally by compartmental analysis on green thalli from Riccia fluitans. Both methods give ratios far too low compared with the thermodynamically possible accumulation of about 10⁴. It is suggested that substantial leakages via different non-electrical pathways prevent equilibrium at steady state, and it is concluded that in such leaky systems the thermodynamic equilibrium condition is not suitable for estimating stoichiometries.