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The primary structure of lactobacillus casei thymidylate synthetase ec 2.1.1.45 part 1 the isolation of cyanogen bromide peptides 1 5 and the complete amino acid sequence of cyanogen bromide 1 2 3 and 5


, : The primary structure of lactobacillus casei thymidylate synthetase ec 2.1.1.45 part 1 the isolation of cyanogen bromide peptides 1 5 and the complete amino acid sequence of cyanogen bromide 1 2 3 and 5. Journal of Biological Chemistry 254(4): 1288-1295

Thymidylate synthetase [EC 2.1.1.45] from L. casei was S-carboxymethylated and degraded by treatment with CNBr. Although the protein contains 6 methionine residues, only 5 CNBr peptides were obtained due to the presence of 1 methionine on the NH2 terminus and another adjacent to a threonine residue which was resistant to cleavage. The peptides were isolated by differential extraction, first with ammonium acetate, then pyridine acetate, and finally the residue was solubilized with 50% acetic acid. Each peptide was further purified to homogeneity by Bio-Gel chromatography. The size of the peptides from the amino to carboxyl end of the enzyme subunit was CNBr 1, 4,100; CNBr 2, 10,300; CNBr 3, 8,100; CNBr 4, 11,800; CNBr 5, 2,200. The sum of the amino acid residues of the peptides is equal to the sum of the residues in an enzyme subunit, indicating that all of the CNBr peptides were isolated. The CNBr-resistant methionine was located in CNBr 2 and the 5-fluoro-2'-deoxyuridine 5'-monophosphate binding site in CNBr 4. The holoenzyme MW, based on the residue weights of the amino acids in the 2 equivalent subunits, is equal to 73,176. The complete sequence of each of the CNBr peptides, except for CNBr 4, is described.

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Maley, G.F.; Bellisario, R.L.; Guarino, D.U.; Maley, F., 1979: The primary structure of Lactobacillus casei thymidylate synthetase. I. The isolation of cyanogen bromide peptides 1 through 5 and the complete amino acid sequence of CNBr 1, 2, 3, and 5. Thymidylate synthetase from Lactobacillus casei was S-carboxymethylated and degraded by treatment with cyanogen bromide. Although the protein contains 6 methionine residues, only 5 cyanogen bromide peptides were obtained due to the presence of 1 m...

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