Biopolym. Cell. 1986; 2(3):129-135, 159.
Structure and Function of Biopolymers
Primary structure of the bacterial formate dehydrogenase determination of N-terminal amino acid sequence and isolation of cysteine-containing peptides
1Ustinnikova T. B., 1Popov V. O., 2, 3Egorov A. M., 3Egorov Ts. A.
  1. A. N. Bakh Institute of Biochemistry, Academy of Sciences of the USSR
    Moscow, USSR
  2. N. I. Vavilov Institute of General Genetics, Academy of Sciences of the USSR
    Moscow, USSR
  3. M. V. Lomonosov State University
    Moscow, USSR

Abstract

NAD-dependent formate dehydrogenase from methylotrophic bacteria of Pseudotnonas sp101 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 cysteine residue responsible for the catalytic enzyme activity was located. A method for the determination of the N-terminal amino acids after fragmentation of the immobilized protein was developed. The N-terminal amino acid sequence of the formate dehydrogenase immobilized via its cysteine residues on a controlled porous glass was determined.

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