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PMID: 4824206 Published · ppublish English Journal Article

Triose phosphate isomerase from the coelacanth. An approach to the rapid determination of an amino acid sequence with small amounts of material.

The Biochemical journal ·Vol. 137 ·No. 2 ·1974-02-00 ·Pages 185-97

Kolb E, Harris JI, Bridgen J

Abstract

The preparation and purification of cyanogen bromide fragments from [(14)C]carboxymethylated coelacanth triose phosphate isomerase is presented. The automated sequencing of these fragments, the lysine-blocked tryptic peptides derived from them, and also of the intact protein, is described. Combination with results from manual sequence analysis has given the 247-residue amino acid sequence of coelacanth triose phosphate isomerase in 4 months, by using 100mg of enzyme. (Two small adjacent peptides were placed by homology with the rabbit enzyme.) Comparison of this sequence with that of the rabbit muscle enzyme shows that 207 (84%) of the residues are identical. This slow rate of evolutionary change (corresponding to two amino acid substitutions per 100 residues per 100 million years) is similar to that found for glyceraldehyde 3-phosphate dehydrogenase. The reliability of sequence information obtained by automated methods is discussed.

MeSH Terms
Amino Acid Sequence Amino Acids/analysis Animals Biological Evolution Carbohydrate Epimerases/analysis Carbon Radioisotopes Chromatography, DEAE-Cellulose Chromatography, Gel Cyanogen Bromide Electrophoresis, Polyacrylamide Gel Evaluation Studies as Topic Fishes Glyceraldehyde-3-Phosphate Dehydrogenases Iodoacetates Methods Microchemistry Muscles Organophosphorus Compounds Peptide Fragments/analysis Rabbits Species Specificity Spectrophotometry, Ultraviolet Thermolysin Trioses Trypsin
Chemicals
Amino Acids Carbon Radioisotopes Iodoacetates Organophosphorus Compounds Peptide Fragments Trioses Glyceraldehyde-3-Phosphate Dehydrogenases Trypsin Thermolysin Carbohydrate Epimerases Cyanogen Bromide
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kolb E
Harris J I
Bridgen J
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29 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1370185
Published
1974-02-00
Pages
185-97
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1166104
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1370185 PMC Full Text View journal details
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