PMID: 8818213Jan 1, 1996

Mechanism of ligand-protein interaction in plant seed thiamine-binding proteins. Preliminary chemical identification of amino acid residues essential for thiamine binding to the buckwheat-seed protein

Maria Rapala-Kozik, A Kozik


Thiamine-binding protein, isolated from buckwheat seeds, was chemically modified in an attempt to identify amino acid residues involved in protein-thiamine interaction. No evidence was found in support of specific roles of arginine residues, sulfhydryl groups, amino groups and tyrosine residues. Under carefully controlled reaction conditions (Tris pH 5-6), the modification with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide caused a complete loss of thiamine-binding capacity. Thus, the carboxyl groups seemed to be essential for binding, possibly for ionic interaction with protein-bound thiamine cation. A selective modification of histidine residues using diethylpyrocarbonate correlated with a loss of thiamine-binding capacity; the modification and the loss of binding capacity could be reversed with hydroxylamine; some ligand-protection against modification was observed. From Tsou analysis of diethylpyrocarbonate modification and resulting loss of thiamine-binding it was suggested that 1-2 of 20 histidine residues of the protein were essential for thiamine binding. The essential histidine(s) might be present in the binding site and possibly were involved in hydrogen bonding(s) with protein-bound thiamine molecule.


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Sep 12, 2019·Plant Signaling & Behavior·Gerry Aplang JanaMahmoud W Yaish


Mar 27, 1979·Biochimica Et Biophysica Acta·A Iwashima, H Nishimura
Mar 15, 1979·Biochemical and Biophysical Research Communications·A L George, C L Borders
Jan 1, 1977·Journal of Supramolecular Structure·G B HendersonF M Huennekens
Sep 21, 1971·Biochimica Et Biophysica Acta·T Nishimune, R Hayashi
Jul 14, 1971·Journal of the American Chemical Society·M H O'Leary, G A Samberg
Sep 1, 1980·Analytical Biochemistry·P J Munson, D Rodbard
Feb 1, 1984·Journal of Nutritional Science and Vitaminology·H NishimuraA Iwashima

Related Concepts

Thiamine Mononitrate
Structure-Activity Relationship
Buckwheat preparation
Sulfhydryl Compounds
Protein-Protein Interaction
Binding Protein
Ligand Binding Domain

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