Structure of the spectrin-actin binding site of erythrocyte protein 4.1

The complete primary structure of the functional site of erythrocyte protein 4.1 involved in spectrin-actin associations has been determined. The sequence of this domain, which contains 67 amino acids and has a molecular mass of 8045 daltons, has been obtained by NH2-terminal sequence analysis of an 8-kDa chymotryptic peptide, three endoproteinase lysine C-cleaved peptides and two peptides obtained by Staphylococcus aureus protease V8 cleavage. All peptides including the 8-kDa domain peptide were purified by reverse-phase high performance liquid chromatography. Antibodies against two different synthetic peptides of the 8-kDa domain are able to inhibit the association between protein 4.1, spectrin, and F-actin, corroborating that the 8-kDa domain is responsible for the formation of a ternary complex. A computer search of the 8-kDa sequence with the National Biomedical Research Foundation database did not detect any significant homologies to known sequences. Protein 4.1 is not related to any known proteins and may represent a new protein superfamily.     

Monosomy 21: a new case confirmed by in situ hybridization

Summary A new case of total monosomy 21 in a newborn is described. The diagnosis was first made using the cytogenetic data; it was then confirmed by the dosage of copper-superoxide dismutase (SOD-1) which showed a 50% decrease. In situ hybridization with a probe previously assigned to chromosome 21 was used to rule out the possibility of a partial monosomy with an unbalanced reciprocal translocation.     

Development of gastric sensitivity to histamine in the rat

Sensitivity of the developing rat stomach to histamine (HA) was examined on isolated gastric mucosae of rats of various ages from the fetal to adult periods. Spontaneous acid secretion in mu eq/h.cm2 occurred at all the ages studied, at a basal rate of 0.45 +/- 0.07 in fetuses to 0.22 +/- 0.03 (day 5), 0.11 +/- 0.04 (day 10), 0.12 +/- 0.04 (day 12), 0.22 +/- 0.08 (day 16) and 0.33 +/- 0.04 (adults). In the fetal rats as in the adults, marked responses to respectively 10(-5) and 10(-4) M HA were demonstrated. The H2-receptor antagonist cimetidine diminished HA-induced secretion by 66 and 57% in fetuses and adults respectively. Between these two stages (from days 5 to 12), basal secretion and the response to HA dropped significantly. On day 21 of gestation, as well as on the critical days 5 and 12 after parturition, db-cAMP (10(-4) M) caused maximal stimulation of acid secretion. These results indicate that the development of responsiveness to HA in the rat is biphasic. They suggest that after birth, the H2-receptor adenylate cyclase system undergoes major modifications which might lead to the complete lack of responsiveness to HA by day 12.     

Influence of arm position on measurement of blood pressure.

A series of measurements of blood pressure in normotensive and hypertensive subjects showed that measurements made with a sphygmomanometer with the arm dependent by the side were consistently higher than those made with the arm horizontal at heart level. The mean difference in a group of 90 hypertensive outpatients was 11/12 mm Hg. Failure to appreciate the importance of arm position may lead to erroneous measurements of blood pressure. This has important implications for clinical practice and research.     

The stimulus-secretion coupling of amino acid-induced insulin release metabolic interaction L-asparagine and L-leucine in pancreatic islets

1. Because L-asparagine augments insulin release evoked by L-leucine, the metabolism of these two amino acids was investigated in rat pancreatic islets. 2. L-Leucine inhibited the uptake and deamidation of L-asparagine, but failed to exert any obvious primary effect upon the further catabolism of aspartate derived from exogenous asparagine. 3. L-Asparagine augmented the oxidation of L-leucine, and effect possibly attributable to activaion of 2-ketoisocaproate dehydrogenase. 4. The association of L-asparagine and L-leucine exerted a sparing action on the utilization of endogenous amino acids, so that the integrated rate of nutrients oxidation was virtually identical in the sole presence of L-leucine and simultaneous presence of L-asparagine and L-leucine, respectively. 5. It is proposed that the enhancing action of L-asparagine upon insulin release evoked by L-leucine is attributable to an increased generation rate of cytosolic NADPH rather than any increase in nutrients oxidation.     

Sulphydryl and Disulphide Levels in Protein Fractions from Hydrated and Dry Leaves of Resurrection Plants

Significant changes in sulphydryl (‘SH’) and disulphide(‘SS’) levels during air-drying in leaves of ‘resurrection’plants (whose protoplasm survives dehydration) stemmed mainlyfrom protein turnover effects. No significant changes were foundin the SH, SS levels in leaves of the desiccation sensitivespecies Sporobolus pyramidalis following air-drying. The three tolerant species studied differed in the directionof change. Some data were consistent with Levitt's SH, SS hypothesis:increases in protein-SS levels in Sporobolus stapfianus (desiccationtolerant) were consistent with a stabilization of new proteinby SS bonds; lower reactivity of protein-SH in the tolerantspecies Talbotia elegans (which on the other hand has decreasedprotein-SS) is consistent with a second mechanism of decreasingprotein denaturation proposed in Levitt's hypothesis. Evidence of some conversion of SH to SS in the soluble proteinsof Xerophyta viscosa (a tolerant species) would on Levitt'shypothesis indicate an injurious process. Some degree of proteindenaturation might be indicated by partial inactivation of thesoluble enzyme ribulose bisphosphate carboxylase in this species,and loss of some soluble isoenzymes (peroxidase and alkalinephosphatase). An apparent lack of SH conversion to SS in thesensitive species Sporobolus pyramidalis was not consistentwith the SH, SS hypothesis. Resurrection plants, Sporobolus pyramidalis, Sporobolus stapfianus, Talbotia elegans, Xerophyta viscosa, drought resistance, desiccation tolerance, protein turnover, sulphydryl groups     

Genes for the biosynthesis of spinosyns: applications for yield improvement in Saccharopolyspora spinosa

Spinosyns A and D are the active ingredients in an insect control agent produced by fermentation of Saccharopolyspora spinosa. Spinosyns are macrolides with a 21-carbon, tetracyclic lactone backbone to which the deoxysugars forosamine and tri-O-methylrhamnose are attached. The spinosyn biosynthesis genes, except for the rhamnose genes, are located in a cluster that spans 74 kb of the S. spinosa genome. DNA sequence analysis, targeted gene disruptions and bioconversion studies identified five large genes encoding type I polyketide synthase subunits, and 14 genes involved in sugar biosynthesis, sugar attachment to the polyketide or cross-bridging of the polyketide. Four rhamnose biosynthetic genes, two of which are also necessary for forosamine biosynthesis, are located outside the spinosyn gene cluster. Duplication of the spinosyn genes linked to the polyketide synthase genes stimulated the final step in the biosynthesis — the conversion of the forosamine-less pseudoaglycones to endproducts. Duplication of genes involved in the early steps of deoxysugar biosynthesis increased spinosyn yield significantly. Journal of Industrial Microbiology & Biotechnology (2001) 27, 399–402. Received 31 May 2001/ Accepted in revised form 09 July 2001