Preparation and characterization of fragments from the N-terminal end of bovine serum albumin under native conditions.
The domain I of BSA, containing residues 1-183 of the protein sequence, was isolated by CNBr treatment. It was further reductively cleaved into two subfragments, N1 and N2, in 8 M urea; the subfragments were regenerated in GSH and GSSG. The fragment N and subfragments N1 and N2 were found to be homogeneous with respect to size and charge. Results for amino acid composition, N-terminal amino acid sequence, thiol groups and M(r) suggested that the fragments N1 and N2 contain residues 88-183 and 1-87 of the intact BSA respectively. Optical studies, intrinsic-viscosity measurements, gel-filtration data and derived hydrodynamic parameters, taken together with the results on proteolytic digestion, showed that fragment N, as well as its subfragments N1 and N2, exist in compact and globular conformation and that the conformation of N2 fragment is more compact than that of the N1 fragment.
Saeed
T
T
Department of Biochemistry, J.N.M.C., Aligarh, India.
Salahuddin
A
A
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002918:Chymotrypsin / Q000378:metabolism; D003429:Cross Reactions; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D005978:Glutathione / Q000031:analogs & derivatives; D019803:Glutathione Disulfide; D007700:Kinetics; D008623:Mercaptoethanol; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000032:analysis; D014357:Trypsin / Q000378:metabolism; D014508:Urea; D014783:Viscosity
D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; D012710:Serum Albumin, Bovine; D008623:Mercaptoethanol; D014508:Urea; D002918:Chymotrypsin; D014357:Trypsin; D005978:Glutathione; D003488:Cyanogen Bromide; D019803:Glutathione Disulfide