The Amino Acid Sequence of Putidaredoxin, an Iron-Sulfur Protein from Pseudomonas putida

Masaru Tanaka,Mitsuru Haniu,Kerry T Yasunobu

doi:10.1016/s0021-9258(19)42529-5

Masaru Tanaka, Mitsuru Haniu + Show 1 more

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https://doi.org/10.1016/s0021-9258(19)42529-5

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Abstract

Abstract The iron-sulfur protein putidaredoxin from Pseudomonas putida contains 106 amino acids arranged in a single peptide chain. The complete amino acid sequence has been determined. The molecular weight including 2 atoms each of iron and sulfur is 11,594. The molecule contains 6 cysteine residues. It is suggested that positions 39, 45, 48, and 85 (or 86) may be chelated to the iron in the iron sulfide reactive center.

Highlights

NH%-terminal sequences of the peptides and protein samples were determined by the manual Edman degradation procedure
COOH-terminal amino acids were determined by use of carboxypeptidase A or B or by a combination of the two enzymes, as described by Ambler [15]
The results are in agreement with the number of residues obtained from the sequence studies, but differ slightly from the amino acid composition reported by Tsai et al if the aspart.ate content is corrected to 13 residues, the amino acid composition of putidaredoxin reported by Tsai et al [5] is identical with the one reported in this paper

Summary

Methods

Am&o Acid Composilion-The amino acid compositions of the peptides and proteins were determined on acid hydrolysates of the samples on a Beckman-Spinco model 120C automatic amino acid analyzer as described by Spackman et al [11]. NH%-terminal sequences of the peptides and protein samples were determined by the manual Edman degradation procedure. The PTH2 derivatives were identified by gas chromatography in a Beckman GC-45 gas chromatograph as described by Pisano and Bronzert [12], by thin layer chromatography as described by Edman and Begg [13], and by amino acid analyses after acid hydrolysis [14]. The. COOH-terminal amino acids were determined by use of carboxypeptidase A or B or by a combination of the two enzymes, as described by Ambler [15]. Hydrazinolysis was performed by the procedure of Bradbury [16] to complement the COOH-terminal amino acid analyses

Results

Discussion

Conclusion