Respiratory properties of cysts and vegetative cells of Azotobacter vinelandii

Abstract The respiratory activity of cysts of Azotobacter vinelandii has been compared with that of vegetative cells. Whole cysts had a much reduced respiratory activity which was less sensitive to KCN. Substrate oxidation rates by membrane preparations from cysts were reduced approximately 10-fold and sensitivity to KCN was decreased by a similar factor. Difference spectra of cyst membranes revealed changes in cytochrome content. Cytochrome oxidase d was apparently absent, cytochrome a ₁ levels were approximately halved whilst those of cytochrome oxidase o were almost doubled. Cytochromes of the b and c -type were present in similar amounts to those in vegetative cells.     

A centralized model for creating shared,standardized, microsatellite data that simplifies inter-laboratory collaboration

We demonstrate an efficient model for standardizing microsatellite DNA data among laboratories studying Oncorhynchus mykiss. Eight laboratories standardized 13 microsatellite loci following allele nomenclature of a central laboratory (average inter-laboratory genotyping concordance >98%). Following this central model, we have currently standardized 298 alleles from throughout the species native range. Although we focus here on O. mykiss, our experiences and recommendation apply equally to other broadly distributed species that may benefit from multi-laboratory collaborative data collection.     

Engineering Strontium Binding Affinity in an EF-hand Motif: A Quantum Chemical and Molecular Dynamics Study

Abstract

Proteins with the ability to specifically bind strontium would potentially be of great use in the field of nuclear waste management. Unfortunately, no such peptides or proteins are known—indeed, it is uncertain whether they exist under natural conditions due to low environmental concentrations of strontium. To investigate the possibility of devising such molecules, one of us (CV), in a previous experimental study [J. Biol. Inorg. Chem. 8, 33440 (2003)], proposed starting from an EF-hand motif of the protein calmodulin and mutating some residues to change the motif's specificity for calcium into one for strontium. In this paper, which represents a theoretical complement to the experimental work, we analyzed small-molecule crystallographic structures and performed quantum chemical calculations to identify possible mutations. We then constructed seven mutant sequences of the EF-hand motif and analyzed their dynamical and binding behaviors using molecular dynamics simulations and free-energy calculations (using the MM/PBSA method). As a result of these analyzes we were able to isolate some characteristics that could lead to mutant peptides with enhanced strontium affinity.     

Immunoglobulin (GM and KM) allotypes and relation to population history in native peoples of British Columbia: Haida and Bella Coola

Differences in the frequencies of GM haplotypes among native peoples of the Americas support the hypothesis that there were three distinct waves of migration from northeast Asia into the Americas: Paleo-Indian, Na-Dene, and Inuit (Eskimo)-Aleut (Salzano and Steinberg: Am. J. Hum. Genet. 17:273-279, 1965; Sukernik and Osipova: Hum. Genet. 61:148-153, 1982; Williams et al.:Am. J. Phys. Anthropol. 66:1-19, 1985; Szathmary: In R Kirk and E Szathmary (eds): Out of Asia: Peopling of the Americas and the Pacific. Canberra: The Journal of Pacific History, Canberra Australian National University, pp. 79-104, 1985). We studied GM allotypes in two linguistically unique populations of Canadian west coast native peoples, the Haida and the Bella Coola, and compared them to GM frequencies in populations that are supposed descendants of the three migrations, in order to investigate the possible genetic relationships of these British Columbia (BC) groups to other native populations. We also estimated the amount of European admixture from the frequency of the Caucasian haplotype, Gm3;5. Results showed that the frequencies in both BC populations of the three common native haplotypes (Gm1,17;21, Gm1,2,17;21, and Gm1,17;15,16), were intermediate between the frequencies in supposed descendants of Paleo-Indian and Na-Dene. These genetic findings are consistent with the controversial hypothesis of archeologist C. Borden (Science 203:963-971, 1979) that, following deglaciation about 13,000 years ago, British Columbia was repopulated by peoples from the north (?Na-Dene) and by culturally distinct peoples from the south (?Paleo-Indian). Caucasian admixture estimates suggested that the Haida and Bella Coola have also experienced moderate amounts (12-20%) of genetic input from European-originating peoples.(ABSTRACT TRUNCATED AT 250 WORDS)     

Specific Lymphocyte Sensitization in Cancer: Is There a Common Antigen in Human Malignant Neoplasia?

From human cancer tissue a basic protein can be extracted by the method which yields encephalitogenic factor when applied to human brain. This tumour basic protein (obtained from several different neoplasms) acts as an antigen in the cytopherometric test for malignant neoplasia and in general gives higher results than does brain basic protein. The reverse is true when degenerative disease of the nervous system is studied. The basic protein extractable from brain and from tumours thus has some degree of specificity probably referable to its amino-acid sequence.     

Identification and purification of calcium ion dependent modulators of actin polymerization from bovine thyroid

We describe the purification of Ca2+-dependent actin modulator proteins from bovine thyroid using DNase I affinity chromatography and diethylaminoethylcellulose chromatography. The 40K actin modulator has been purified to 98% homogeneity. It is a single polypeptide chain with a molecular weight of approximately 40 000 and an isoelectric point of 8.1. Its amino acid composition is different from previously described actin-associated proteins and thyroid actin. On the basis of the centrifugation assay and the DNase I inhibition assay, the actin complexed with the 40K protein is G-actin in its conformation rather than F-actin oligomers. Substoichiometric concentrations of the 40K protein rapidly inhibit actin polymerization in the presence of physiological concentrations of Ca2+ and Mg2+. An 80K actin modulator also has been purified to 98% homogeneity. It is a single polypeptide chain with a molecular weight of approximately 80 000 and an isoelectric point of 6.35-7.0. Its amino acid composition is different from those of villin, gelsolin, and leukocyte actin polymerization inhibitor. On the basis of the DNase inhibition assay and the centrifugation assay, the nonprecipitable actin associated with the 80K protein was F-actin in its conformation. The 80K protein acts very efficiently as a Ca2+-dependent nucleator for actin assembly and reduces its viscosity. In addition to the 40K and 80K actin modulators, 91K and 95K actin-associated proteins were partially purified. The 91K-95K fraction has similar activity to the 80K protein regarding precipitation of F-actin. The 125I-G-actin polyacrylamide gel overlay technique [Snabes, M. C., Boyd, A.E., & Bryan, J. (1981) J. Cell Biol. 90, 809-812] revealed that both the 91K and 95K proteins bind 125I-actin after sodium dodecyl sulfate (NaDodSO4) electrophoresis while the 80K and 40K proteins do not. Thyroid 91K protein comigrated with a human platelet 91K actin binding protein on NaDodSO4 gels and may be similar to macrophage gelsolin. The 95K protein may be similar to villin, the intestinal cytoskeletal protein.