Identification of zinc-binding sites of proteins: zinc binds to the amino-terminal region of tubulin.
The discovery that certain proteins may require zinc for their activity, and the fact that several of them cannot be purified in large amounts, has led us to develop a rapid, sensitive method to detect these proteins in samples. This method is based on the fractionation of the proteins by gel electrophoresis, blotting onto nitrocellulose paper, and overlaying with 65Zn. We have tested the procedure with well-characterized zinc-binding proteins. In the case of tubulin, we have used this method to localize its zinc-binding site. It was found that zinc binds to the first 150 amino acids of both alpha- and beta-tubulin subunits.
Serrano
L
L
Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma, Madrid, Spain.
Dominguez
J E
JE
Avila
J
J
D001665:Binding Sites; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002132:Calcium Radioisotopes; D002352:Carrier Proteins / Q000032:analysis*; D003035:Cobalt / Q000378:metabolism; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010084:Oxidation-Reduction; D014404:Tubulin / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*; D015037:Zinc Radioisotopes
D002132:Calcium Radioisotopes; D002352:Carrier Proteins; D014404:Tubulin; D015037:Zinc Radioisotopes; C017800:zinc-binding protein; D003035:Cobalt; D003101:Collodion; D015032:Zinc; D002118:Calcium