Biopolymers and cell. 1986. Volume 2. № 3. 129 - 135
T. B. Ustinnikova, V. 0. Popou, A. M. Egorou, Ts. A. Egorov
PRIMARY STRUCTURE OF THE BACTERIAL FORMATE DEHYDROGENASE DETERMINATION OF N-TERMINAL AMINO ACID SEQUENCE AND ISOLATION OF CYSTEINE-CONTAINING PEPTIDES
Summary
NAD-dependent formate dehydrogenase from methylotrophic bacteria of Pseudotnonas splOl was immobilized on thiopropyl-Sepharose 6B and CPG/Thiol activated by 2,2'-di-pyridyl disulphide via the thiol-disulphide exchange reaction. A procedure for the tryptic digestion of the immobilized enzyme was optimized. Cysteine-containing tryptic peptides were isolated and partially characterized. The peptide which contained the essential cystei-ne residue responsible for the catalytic enzyme activity was located. A method for the de¬termination of the N-terminal amino acids after fragmentation of the immobilized pro¬tein was developed. The N-terminal amino acid sequence of the formate dehydrogenase immobilized via its cysteine residues on a controlled porous glass was determined.