Highly basic cyanogen bromide peptides from the proteins of turnip rosette virus and the Prunus strain of tomato bushy stunt virus

1986 ◽  
Vol 64 (12) ◽  
pp. 3043-3046
Author(s):  
J. H. Tremaine ◽  
W. P. Ronald
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Mosaic Virus ◽  
Acid Composition ◽  
Cyanogen Bromide ◽  
Sodium Dodecyl ◽  
Tomato Bushy Stunt Virus ◽  
Basic Peptide ◽  
Amino Terminal ◽  
Basic Peptides

The proteins of turnip rosette virus (TRosV) a sobemovirus, and the Prunus strain of tomato bushy stunt virus (TBSV-P) were cleaved with cyanogen bromide, and highly basic peptides were isolated by ion-exchange chromatography. The amino acid composition and sodium dodecyl sulphate – polyacrylamide gel electrophoresis indicated the basic peptide from TRosV had 40 amino acid residues, including 11 basic residues. This peptide had a composition similar to basic peptides isolated from the sobemoviruses, sowbane mosaic virus, and two strains of southern bean mosaic virus. The amino acid composition and size of the basic peptide from TBSV-P were similar to those found for the amino terminal sequence of the type strain of TBSV.

scholarly journals Separation and partial characterization of guinea-pig caseins

1978 ◽  
Vol 173 (2) ◽  
pp. 633-641 ◽  
Author(s):  
R K Craig ◽  
D McIlreavy ◽  
R L Hall
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Guinea Pig ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Exchange Chromatography

1. Guinea-pig caseins A, B and C were purified free of each other by a combination of ion-exchange chromatography and gel filtration. 2. Determination of the amino acid composition showed all three caseins to contain a high proportion of proline and glutamic acid, but no cysteine. This apart, the amino acid composition of the three caseins was markedly different, though calculated divergence values suggest that some homology may exist between caseins A and B. Molecular-weight estimates based on amino acid composition were in good agreement with those based on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 3. N-Terminal analysis showed lysine, methionine and lysine to be the N-terminal residues of caseins A, B and C respectively. 4. Two-dimensional separation of tryptic digests revealed a distinctive pattern for each casein. 5. All caseins were shown to be phosphoproteins. The casein C preparation also contained significant amounts of sialic acid, neutral and amino sugars. 6. The results suggest that each casein represents a separate gene product, and that the low-molecular-weight proteins are not the result of a post-translational cleavage of the largest. All were distinctly different from the whey protein alpha-lactalbumin.

Amino acid composition and the amino terminal end groups of the α- and β-chains of four Lemur hemoglobins

1965 ◽  
Vol 109 (3) ◽  
pp. 571-578 ◽  
Author(s):  
Ralph A. Bradshaw ◽  
Larry A. Rogers ◽  
Robert L. Hill ◽  
John Buettner-Janusch
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Amino Terminal ◽  
End Groups

scholarly journals Studies on the subunits of human myeloperoxidase

1984 ◽  
Vol 222 (3) ◽  
pp. 701-709 ◽  
Author(s):  
R L Olsen ◽  
C Little
Keyword(s):  
Heat Treatment ◽  
Amino Acid ◽  
Molecular Mass ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Additional Species ◽  
Main Protein

The subunit composition of human myeloperoxidase was studied with the use of sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and gel filtration. The subunit pattern observed depended on the manner in which the enzyme was treated before analysis. Reduction before heat treatment in detergent led to two main protein species (Mr 57 000 and 10 500), whereas reduction during or after heat treatment yielded an additional species of Mr 39 000. Heating without any reductive pretreatment yielded the 39 000-Mr form as the major electrophoretic species. Carbohydrate staining showed large amounts of sugar on the 57 000-Mr species and little on the 10 500-Mr form. Significant amounts of haem were associated with this latter subunit. Haem also seemed to be associated with the 57 000-Mr form but not with the 39 000-Mr one. These three subunit forms were isolated and their amino acid composition analysed. The 57 000-Mr and 39 000-Mr forms had very similar amino acid composition and yielded an apparently identical collection of fragments on incubation with CNBr. Once separated, the subunits could not be interconverted. Generally, minor amounts of other molecular-mass forms were observed. The nature of the various molecular-mass forms originating from myeloperoxidase is discussed.

Chemical and serological properties of a cyanogen bromide peptide of southern bean mosaic virus protein

1980 ◽  
Vol 26 (12) ◽  
pp. 1450-1459 ◽  
Author(s):  
J. H. Tremaine ◽  
W. P. Ronald ◽  
E. M. Kelly
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Mosaic Virus ◽  
Cyanogen Bromide ◽  
Tomato Bushy Stunt Virus ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Residues ◽  
Southern Bean Mosaic Virus ◽  
Gel Diffusion

Southern bean mosaic virus (SBMV) protein was cleaved with cyanogen bromide and a highly basic peptide, CB-1, was isolated by ion exclusion and ion-exchange chromatography. Twelve peptides were separated from a tryptic digest of CB-1 by ion-exchange chromatography and the composition of these peptides was similar to that of peptides released from EDTA-swollen virus particles by limited tryptic digestion. The composition and N-termini of the tryptic peptides indicated CB-1 was from the N-terminus of SBMV protein and contained 48 amino acid residues. The CB-1 peptide moved rapidly to the cathode in polyacrylamide gel electrophoresis at pH 3.9 and contained nine arginine residues, three lysine residues, and no acidic amino acid residues. It was shown to interact with purified viral RNA, sodium dextran sulfate, and calf thymus DNA.Antiserum to sodium dodecyl sulfate (SDS)-dissociated virus gave a reaction of partial identity between the CB-1 peptide and the SDS-dissociated virus in SDS gel diffusion tests. The CB-1 peptide did not react with antiserum to SDS-dissociated, trypsin-treated virus. Gel diffusion tests conducted in saline agar gels between trypsin-treated virus and SBMV, with SBMV antiserum, did not show differences in their serological properties. Antiserum to the CB-1 peptide conjugated to tomato bushy stunt virus reacted with SBMV but SBMV antiserum did not react with CB-1 or the CB-1-tomato bushy stunt virus conjugate.

scholarly journals Localization and Characterization of the Cleavage-Associated Neoantigen in the Edomain of Fibrinogen

1977 ◽  
Author(s):  
E. F. Plow ◽  
T. S. Edgington
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Conformational Changes ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Terminal ◽  
Exclusion Chromatography

Plasmic cleavage of fibrinogen to generate fragment X partially exposes a specific cryptic molecular site, fg-Eneo. This site in the E domain of the molecule is further exposed during subsequent cleavage. We now report on localization of this site which provides an incisive marker for the structural and conformational changes associated with plasmic cleavage of fibrinogen. Fg-Eneo was stable to reduction and alkylation and the chains of the E fragment were separated by ion exchange chromatography on DEAE-cellulose. An active component was obtained and subjected to molecular exclusion chromatography on Sephadex G-50 to insure removal of intact fg-E. A fg-Eneo positive chain was recovered and identified as Eγ with respect to amino-terminal tyrosine, amino acid composition, and immunochemical analysis. The fg-Eneo site was stable to tryptic degradation, and tryptic peptides were prepared and separated by multiple molecular exclusion chromatographic steps. Final separation of two peptides of similar size was achieved on the basis of carbohydrate content by affinity chromatography on Concanavalin A. Only the active peptide was bound by the lectin. Purity and identification of the active tryptic peptide as γ36–53 was established by amino acid composition and sequence. These results establish that this region of the γ chain of fibrinogen is not present at the hydrated surface of the native molecule but that, in association with plasmic cleavage and conformational changes, this site is progressively exposed and provides a dynamic marker of the cleavage sequence.

Changes in renal glomerular basement membrane with age and nephritis

1977 ◽  
Vol 55 (12) ◽  
pp. 1197-1206 ◽  
Author(s):  
N. Kalant ◽  
S. Satomi ◽  
R. White ◽  
E. Tel
Keyword(s):  
Amino Acid ◽  
Basement Membrane ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Glomerular Basement Membrane ◽  
Disulfide Bonds ◽  
Sodium Dodecyl ◽  
Collagen Content ◽  
Insoluble Residue ◽  
Adult Rats

Glomerular basement membrane was obtained from normal, young and old adult rats and from animals with antiserum nephritis, daunomycin nephrosis, and lathyrism. With increased age there was an increase in the collagen content of whole glomeruli and of the basement membrane. About 50% of the membrane protein was solubilized in 1% sodium dodecyl sulfate, and a further 35% was solubilized by reduction and alkylation. Inhibition of formation of collagen cross-links by induction of lathyrism did not affect membrane solubility. Preparative disc gel electrophoresis permitted separation of a number of components of different composition; with decreasing molecular weight the collagen content declined from almost 100% to 0%.In antiserum nephritis, there was an increase in the noncollagen components of the membrane and marked alterations in amino acid composition, both of whole membrane and of electrophoretically separated components. In daunomycin nephrosis, the amino acid composition was similar to that of antiserum nephritis. The solubility of membrane from nephritic rats was normal. The composition of the insoluble residue was similar for all membrane preparations and resembled pure collagen. It is suggested that the presence of abnormal noncollagen proteins, associated with the insoluble collagen core by hydrophobic and disulfide bonds, as in antiserum nephritis, is associated with increased membrane permeability leading to proteinuria.

scholarly journals Purification and properties of a proteolytic enzyme from the cercariae of the human trematode parasite Schistosoma mansoni

1982 ◽  
Vol 201 (1) ◽  
pp. 137-144 ◽  
Author(s):  
W J Landsperger ◽  
M A Stirewalt ◽  
M H Dresden
Keyword(s):  
Amino Acid ◽  
Schistosoma Mansoni ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Isoelectric Point ◽  
Proteolytic Enzymes ◽  
Sodium Dodecyl ◽  
Skin Penetration ◽  
Cathepsin G ◽  
Trematode Parasite

Skin penetration by the cercarial stage of the human trematode parasite Schistosoma mansoni is mediated by the secretion of proteolytic enzymes able to digest components of mammalian connective tissues. In the present study the purification of these proteinases from cercarial homogenates is reported. The major proteinase species has a mol. wt. of approx. 25 000 and exists in monomeric form as determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. This proteinase has an isoelectric point of 6.0. Studies presented here, with a variety of substrates and inhibitors, confirm previous claims that these proteinases belong to the serine class, and, in addition, suggest that they resemble the vertebrate chymotrypsins rather than trypsins or elastases. However, the amino acid composition of the cercarial proteinase differs significantly from bovine chymotrypsin and from the human leucocyte chymotrypsin-like cathepsin G. The amino-acid-composition differences between these proteinases are consistent with their differences in isoelectric point. In order to obtain an insight into the role of the proteinase in skin penetration, its activity on cartilage proteoglycan monomers and on the isolated peptide backbone of proteoglycan was studied. The results of the present study indicate that the cercarial enzyme catalyses a limited specific digestion of the peptide core.

scholarly journals Isolation, composition and ordering of cyanogen bromide peptides in human casein

1978 ◽  
Vol 169 (2) ◽  
pp. 337-342 ◽  
Author(s):  
M L Groves ◽  
R Greenberg
Keyword(s):  
Amino Acid ◽  
Human Milk ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Cyanogen Bromide ◽  
Limited Digestion

The major component of the casein fraction of human milk was cleaved by cyanogen bromide, and the composition of the resulting peptides was determined. Casein was also subjected to limited digestion by trypsin, and the amino acid composition of the isolated peptides was established. With this information the peptides were ordered as they occur in the purified protein.

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