Glutathione Reductase from Human Erythrocytes

• DOI: 10.1111/j.1432-1033.1982.tb05780.x
• Publication Date: 2005-03-03T21:30:15Z
• Authors: R. Luise KRAUTH‐S...; Mansoor SALEH; Renate UNTUCHT‐GRAU; R. Heiner SCHIRMER; Rosi BLATTERSPIEL; Emile SCHILTZ
• Keywords: Edman degradation; Complete sequence; Sequence (biology)

Sequence analysis of the NADPH domain (residues 158 – 293) and interface (365 478) was based on 12 CNBr fragments, which were isolated using ion‐exchange chromatography paper methods. Fragments with more than 15 residues digested further trypsin chymotrypsin. The peptides sequenced by automated solid‐phase Edman degradation. All ordered overlapped computerized comparisons a complete sequence guessed from electron density map protein. In case short this alignment confirmed protein fragments resulting incomplete cleavage. domain, residue 197, is involved in an induced‐fit mechanism, identified as tyrosine. structure probably homologous NAD lipoamide dehydrogenase FAD several proteins, but not domains known chainfold other proteins. completes glutathione reductase so that enzyme now atomic detail. numbering scheme chemically determined will be used henceforth crystallo‐ graphic studies also. As inferred data each two identical chains contains 478 amino acid residues, composition being Cys 10 , Asp 21 Asn 17 Thr 31 Ser Glu 29 Gln 11 Pro 24 Gly 43 Ala 42 Val 44 Met He Leu 34 Tyr 13 Phe 14 Lys His 16 Arg l7 Trp 3 . From these M r 2 × 51600 calculated for FAD‐free apoenzyme 52400 holoenzyme.