Search PubMed⌕ Search

Biomedical subjects

G Boileau

Publications and source records attributed to G Boileau.

106 records · Page 6Linked to original sources
PubMed1981

Identification of proteins at the binding site for protein S1 in 70 S ribosomes and 30 S subunits by cross-linking with 2-iminothiolane.

Escherichia coli 30 S ribosomal subunits and 70 S ribosomes were treated with 2-iminothiolane and oxidized to promote the formation of intermolecular disulfide bonds between neighboring proteins. After alkylation of the unreacted sulfhydryl groups, the mixture of ribosomal proteins was extracted with acetic acid and analyzed by 2-dimensional polyacrylamide gel electrophoresis using nonequilibrium pH gradient gel electrophoresis in the first dimension and a sodium dodecyl sulfate gel in the second. The analysis revealed only 3 protein dimers that contained ribosomal protein S1. The identification of the protein partner of protein S1 in each dimer was accomplished by radioiodination of the proteins extracted from the gel, 2-dimensional polyacrylamide/urea gel electrophoresis, and radioautography. The results indicate that proteins S2, S10, and S18 are cross-linked to protein S1. No other cross-links involving S1 were detected.

Binding Sites↗
PubMed1978

Streptomycin-induced conformational changes in the 70-S bacterial ribosome.

Conformational alterations induced by streptomycin in the bacterial ribosome have been investigated using as probes, ethidium bromide, N-[14C]ethylmaleimide and a spin label nitroxide analog of N-ethylmaleimide. 1. The binding of the antibiotic to the ribosome does not affect the reactivity of sulfhydryl groups towards N-ethylmaleimide. 2. The motional freedom of spin labels bound to ribosomal proteins S1 and S18 is increased but it is hardly affected at other labeled sites. This observation suggests that the binding of streptomycin causes a local loosening of the ribosomal structure. 3. Ribosomes are found to bind less ethidium bromide in the presence of streptomycin, which suggests that the binding of streptomycin decreases the degree of organization of ribosomal RNA.

Electron Spin Resonance Spectroscopy↗
PubMed1976

The use of N-succinyl derivatives in the study of amino acids and peptides by mass spectrometry.

The terminal amino group of amino acids and peptides is blocked as the N-succinyl derivative by reaction with succinic anhydride. The product is then converted to the N,O-permethyl derivative in order to increase its volatility for use in mass spectrometry. The permethylated N-succinyl derivative retains the advantages of the permethylated N-acetyl derivative in regard to ease of preparation on a small scale, volatility and the presence of characteristic fragmentation patterns in their mass spectra. However, peaks in the high mass region are more abundant due to loss of CH3O-from the N-succinyl carbomethoxyl group as well as from the C-terminal carbomethoxyl group. Ions characteristic of the sequence and of individual amino acids are observed, and molecular weight can be determined from the relatively abundant ion at [M--CH3O]+ and from the weak molecular ion.

Amino Acids↗
PubMedDate not supplied

Complete sequence of rabbit kidney aminopeptidase N and mRNA localization in rabbit kidney by in situ hybridization.

We have isolated and sequenced a full-length cDNA for rabbit kidney aminopeptidase N (APN). The 3-kilobase cDNA contains 12 nucleotides of the 5' noncoding region, a 2898 nucleotide long open reading frame, and 113 nucleotides of the 3' untranslated region. The open reading frame encodes a type II membrane protein of 966 amino acid residues composed of a 10 residue NH2-terminal cytosolic domain, a 23 residue transmembrane domain, and a large 933 residue ectodomain that contains the active site. Rabbit APN has eight potential N-glycosylation sites and seven cysteine residues, one of which is located in the transmembrane domain. Computer analysis showed that the enzymes from human, rat, and rabbit were highly conserved, except for the stalk region immediately downstream from the transmembrane domain. Using in situ hybridization techniques we showed that in rabbit kidney, APN mRNA is present in proximal tubules but not in glomeruli, which corresponds to the localization of the protein observed by immunohistochemistry. Taken together, our results strongly suggest that the expression of APN in kidney is modulated at mRNA levels and not at translational and (or) posttranslational levels.

Amino Acid Sequence↗
PubMedDate not supplied

Translocation of neutral endopeptidase 24.11 mutants with deletions of the NH2-terminal cytosolic domain.

Rabbit neutral endopeptidase 24.11 (NEP) is a type II membrane protein with a positively charged 27 amino acid residue NH2-terminal cytoplasmic domain, a 20 amino acid residue hydrophobic signal peptide/membrane anchor domain, and a large catalytic COOH-terminal domain exposed on the exoplasmic side of the membrane. To study the role of the cytosolic domain in anchoring NEP in the plasma membrane, we constructed two mutants in which this cytosolic domain was deleted. In the first mutant (NEP delta cyto), a Glu residue was present in NH2-terminus, while a Lys residue was substituted at the same position in the second mutant (NEP delta cyto(K)). To better understand the interaction of these mutants with the rough endoplasmic reticulum membrane, the mutated NEP cDNAs were transcribed and translated in vitro in the presence of microsomal membranes. Our studies showed that deletion of the hydrophillic cytosolic domain affects translocation of the NEP polypeptide chain. Substitution of a positively charged Lys residue for the Glu residue at the NH2-terminus of the deletion mutant only partly restored translocation of the polypeptide chain. Furthermore, carbonate extraction and trypsin digestion of the microsomal membranes indicated that the deletion mutants are inserted in the microsomal membranes as type III membrane proteins with their COOH-terminal domain exposed on the exterior of the microsomes. Thus, efficient translocation is dependent on the presence of a charged cytoplasmic domain.

Amino Acid Sequence↗
PubMedDate not supplied

ACTH/beta-endorphins and ACTH/cortisol ratios as early biological markers in HIV infection.

Three groups of male homosexuals: AIDS (n = 19), HIV seropositive (n = 15), and seronegative partners of seropositive subjects (n = 15), were compared to a heterosexual seronegative control group (n = 13). Twice daily evaluations (8 A.M. and 5 P.M.) of plasma levels of beta-endorphin, ACTH, and cortisol were done by radioimmunoassay. Seropositive subjects and their seronegative partners showed similar levels of neurohormones: 1. An elevation in the ACTH/beta-endorphin ratio in the plasma, (C = 0.69 +/- 0.84, AIDS = 0.44 +/- 0.32, S+ = 0.42 +/- 0.28, and S- = 0.42 +/- 0.5); 2. A loss of normal relationship of the coupling ACTH/total cortisol, (C = 0.00035 +/- 0.00028, AIDS = 0.00042 +/- 0.0034, S+ = 0.00074 +/- 0.00068, and S- = 0.00072 +/- 0.0008. Neuroendocrinological disorders have been observed in HIV-infected subjects and in their seronegative partners. These could be related to their sexual behavior, as well as to the HIV infection. If this last hypothesis is confirmed, the ACTH/beta-endorphin and ACTH/cortisol ratios may be seen as possible early signs of HIV infection.

Adolescent↗
PubMedDate not supplied

Prediction of the left ventricular ejection fraction response to doxorubicin using a multiple linear regression model.

The aim of this study was to develop a model for predicting clinically significant deterioration in the left ventricular ejection fraction due to chronic doxorubicin administration. Twenty-six patients were monitored during their courses of doxorubicin chemotherapy with serial gated equilibrium radionuclide angiography. Multiple linear regression analysis was used to derived the best combination of clinical and radionuclide angiographic predictors of resting left ventricular ejection fraction at any point during the course of chemotherapy. The final model consisted of five variables: left ventricular ejection fraction at the previous monitoring point; cumulative dose of doxorubicin achieved at the previous monitoring point; increment in dose from the previous monitoring point; age of the patient; and time to peak left ventricular emptying at the previous monitoring point. The cumulative dose, the ejection fraction at the previous monitoring point and the final model, respectively, explained 11%, 33% and 53% of the variability in ejection fraction determinations during the 26 patient courses. The final model also forecast a potentially very low resting left ventricular ejection fraction (less than 35%) at the cumulative doses of doxorubicin which provoked serious clinical cardiotoxicity in two patients. A multivariate model is a useful aid in timing discontinuation of doxorubicin prior to the development of a clinically significant deterioration in left ventricular ejection fraction.

Computer Simulation↗