Primary structure of the bacterial formate dehydrogenase determination of N-terminal amino acid sequence and isolation of cysteine-containing peptides
DOI:
https://doi.org/10.7124/bc.0001A8Abstract
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.References
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