Crossing of low-lying electronic levels of high-spin ferrous ion in deoxyhemoglobin and deoxymyoglobin.

The electronic structure of high-spin S = 2 ferrous ion in deoxy forms of hemoglobin and myoglobin is considered in terms of spin Hamiltonian formalism. Spin Hamiltonian parameters of the second order B0(2)(D), B2(2)(E) and, for the first time in the available literature, of fourth order B0(4), B2(4) and B4(4), are calculated for the rhombic symmetry case of Fe2+. The Hamiltonian matrix is diagonalized for several sets of Bq(k) parameters compatible with other experimental data. The low-lying Fe2+ levels exhibit crossings in a high magnetic field, applied along the z-axis perpendicular to the heme plane. The cross-over values of the magnetic field are determined to be Hc1 = 46 kOe and Hc2 = 168 kOe for D = 5.2, E = 0.6 cm-1 (close to the magnetic data of Nakano, N., Otsuka, J. and Tasaki, A. (1972) Biochim. Biophys. Acta 278, 355-371) and with B0(4) = 0.037, B2(4) = 0.005, B4(4) = 0.013 cm-1 and gz = 2.028. Experimental techniques for measurement of the crossing effects are discussed.     

Thiocyanate ion formation in rapeseed meals.

Meal prepared from unheated rapeseed (Brassica napus cv. Zephyr) showed the presence of t,iocyanate ion, while meal from heated seed of the same cultivar did not show detectable amounts. Unheated seed meal on autolysis, and heated seed meal on incubation with thioglucosidase, yielded increased amounts of thiocyanate ion. Various commercial rapeseed meals showed the presence of t,iocyanate ion only after enzyme incubation. Low glucosinolate, cv. Bronowski, and higher glucosinolate, cv. Zephyr on enzymic incubation yielded comparable amounts of thiocyanate ion, suggesting that the precursor responsible in the two varieties was the same and present in similar quantities. No formation of thiocyanate ion was observed on incubation of sinigrin with thioglucosidase. Rats dosed with heated meal, containing intact glucosinolate, showed a slight increase of thiocyanate ion in the urine as compared with control rats dosed with water, while a relatively large increase followed dosing with sinigrin. Rats dosed with meal containing free thiocyanate ion excreted the ingested thiocyanate ion almost quantitatively.     

Changes in calmodulin level and cAMP-dependent protein kinase activity during epididymal maturation of ram spermatozoa

Calmodulin concentration and cAMP-dependent protein kinase activity were simultaneously determined on ram spermatozoa collected by cannulation of successive segments of the epididymal tubule. Epididymal transit was characterized on one hand by an overall decrease in the calmodulin level and on the other by a dramatic rise in the cAMP-dependent protein kinase activity. In contrast to the calmodulin level, the cAMP-dependent protein kinase activity was correlated with the acquisition of flagellar beat. No further alterations in the level of these two proteins could be detected as spermatozoa acquired progressive motility.     

Seraspenide (acetylSDKP): phase I-II trial study of inhibitor of hematopoiesis protects against toxicity of aracytine and ifosfamide monochemotherapies]

Seraspenide, a synthetic tetrapeptide, inhibits cell cycle entry of normal hematopoietic stem cells. In mice it protects hemopoiesis against the damage caused by cytarabine, cyclophosphamide and carboplatin. Seraspenide has been given to 53 cancer patients undergoing monochemotherapy with cytarabine and ifosfamide in a double-blind cross-over randomized study. A significant protection of peripheral blood cells has been observed. Seraspenide has been devoided of toxicity.     

Protein processing as a regulatory mechanism in the synthesis of the photosynthetic antenna in Chloroflexus

Chloroflexus aurantiacus can be induced to shift from respiratory to photosynthetic energy production by introducing light and/or lowering the oxygen concentration of a culture. After induction, cells synthesize bacteriochlorophyll and proteins for the formation of a functional photosynthetic apparatus. Bacteriochlorophyll is detectable within 2 h after induction. Chlorosome polypeptides are detected after 8–12 h. Two proteins, M_r 60,000 and M_r 47,000, are present in both induced and noninduced cells and react specifically with antibodies against chlorosome polypeptides. Immunological data suggest that these proteins (M_r 60,000 and 47,000) are polyproteins which are transcribed and translated in the dark. When cells are exposed to light or low oxygen tension these proteins are processed into functional polypeptides required in the assembly of the chlorosome. The reaction center polypeptide (M_r 26,000) appears to be part of a separate genetic control system.Dedicated to Prof. G. Drews on occasion of his 60th birthday     

Overproduction from a cellulase gene with a high guanosine-plus-cytosine content in Escherichia coli

A recombinant exoglucanase was expressed in Escherichia coli to a level that exceeded 20% of total cellular protein. To obtain this level of overproduction, the exoglucanase gene coding sequence was fused to a synthetic ribosome-binding site, an initiating ATG, and placed under the control of the leftward promoter of bacteriophage lambda contained on the runaway replication plasmid vector pCP3 (E. Remaut, H. Tsao, and W. Fiers, Gene 22:103-113, 1983). With the exception of an inserted asparagine adjacent to the initiating ATG, the highly expressed exoglucanase is identical to the native exoglucanase. The overproduced exoglucanase can be isolated easily in an enriched form as insoluble aggregates, and exoglucanase activity can be recovered by solubilization of the aggregates in 6 M urea or 5 M guanidine hydrochloride. Since the codon usage of the exoglucanase gene is so markedly different from that of E. coli genes, the overproduction of the exoglucanase in E. coli indicates that codon usage may not be a major barrier to heterospecific gene expression in this organism.