Mapping the functional domains of the eukaryotic elongation factor 1 beta gamma.
The functional domains of the eukaryotic elongation factor (EF) 1 beta gamma have been delineated with the use of limited proteolysis, protein microsequencing, gel electrophoresis under non-denaturing conditions and antibodies against EF-1 beta and EF-1 gamma. By means of limited proteolysis, it was possible to obtain large fragments of EF-1 beta. In contrast to amino-terminal fragments, those derived from the carboxy-terminal part of EF-1 beta were still active in enhancing the guanine nucleotide exchange of GDP bound to EF-1 alpha. With the same technique of limited proteolysis, it was possible to isolate a trypsin-resistant core from EF-1 beta gamma containing polypeptide chain fragments derived from both subunits. A polyvalent antiserum against EF-1 beta and two monoclonal antibodies against EF-1 gamma were used to identify the protein fragments in this core. The monoclonal antibodies were shown to recognize different epitopes, one localized on the amino-terminal and another on the carboxy-terminal half of EF-1 gamma. The antiserum against EF-1 beta and one of the monoclonal antibodies (mAb 36E5), which recognized the amino-terminal half of EF-1 gamma, reacted with this trypsin-resistant core. We conclude that the amino-terminal halves of both EF-1 beta and EF-1 gamma are firmly attached to each other, and that the carboxy-terminal part of EF-1 beta interacts with EF-1 alpha.
van Damme
H
H
Department of Medical Biochemistry, Sylvius Laboratory, University of Leiden, The Netherlands.
Amons
R
R
Janssen
G
G
Möller
W
W
D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D006153:Guanosine Diphosphate / Q000378:metabolism; D006868:Hydrolysis; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics* / Q000276:immunology; D012261:Ribonucleoproteins / Q000235:genetics*
D000911:Antibodies, Monoclonal; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D012261:Ribonucleoproteins; D006153:Guanosine Diphosphate