Microsomal glutathione transferase. Primary structure.
The primary structure of rat liver microsomal glutathione transferase has been determined. The 14C-carboxymethylated protein was fragmented with CNBr and proteolytic enzymes. The basis of the analysis was information from sequenator degradations of the intact protein, the largest CNBr fragment, and a large COOH-terminal fragment derived from a digest with Glu-specific staphylococcal protease. Remaining, smaller fragments were analyzed with the manual dimethylaminoazobenzene isothiocyanate method. Pepsin and limited acid hydrolysis were used to obtain peptides to confirm and overlap hydrophobic structures in the COOH-terminal half of the protein where trypsin and chymotrypsin failed to give any cleavage. Combined, these data permit the deduction of a 154-residue amino acid sequence. No evidence for micro-heterogeneity was obtained. The NH2-terminal alanine residue has a free alpha-amino group and the cysteine residue involved in activation of the enzymatic activity by sulfhydryl reagents is at position 49. The protein chain contains three regions with predictions for long beta strand secondary structures (positions 11-26, 103-120, and 131-145). Predictions may be inaccurate in membrane-associated proteins, but two of these regions also affect the three most hydrophobic segments. Thus, residues 11-35 form a long, largely hydrophobic part interrupted by only one charged residue (Lys-25), and residues 81-97 and 114-126 constitute the most hydrophobic segments directly noticeable from the hydrophilicity curve of the protein chain. These special parts of the molecule are of interest in relation to membrane interactions.
D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin / Q000378:metabolism; D003488:Cyanogen Bromide; D010450:Endopeptidases / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D008666:Metalloendopeptidases; D008862:Microsomes, Liver / Q000201:enzymology*; D017830:Octoxynol; D010434:Pepsin A / Q000378:metabolism; D010446:Peptide Fragments; D010447:Peptide Hydrolases / Q000378:metabolism; D011092:Polyethylene Glycols; D051381:Rats; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D011092:Polyethylene Glycols; D017830:Octoxynol; D005982:Glutathione Transferase; D010450:Endopeptidases; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D014357:Trypsin; D010434:Pepsin A; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D003488:Cyanogen Bromide