Omer, C. A., Kral, A. M., Diehl, R. E., Prendergast, G. C., Powers, S., Allen, C. M., Gibbs, J. B., Kohl, N. E. (May 1993) Characterization of recombinant human farnesyl-protein transferase: cloning, expression, farnesyl diphosphate binding, and functional homology with yeast prenyl-protein transferases. Biochemistry, 32 (19). pp. 5167-76. ISSN 0006-2960 (Print)0006-2960
Abstract
We have isolated cDNAs encoding the alpha and beta subunits of human farnesyl-protein transferase (FPTase). The proteins encoded by these two cDNAs are 93-95% identical to the corresponding subunits of bovine and rat FPTase and show regions of homology with proteins encoded by Saccharomyces cerevisiae prenyl-protein transferase genes. Human FPTase expressed in Escherichia coli from a translationally coupled operon had kinetic properties similar to those of FPTase isolated from bovine brain. Examination of farnesyl diphosphate binding indicated that while neither individual subunit was capable of isoprenoid binding, a radiolabeled farnesyl diphosphate analog could be specifically photo-cross-linked to the beta subunit of FPTase holoenzyme. To further analyze subunit structure-function and to detect functional similarities with yeast prenyl-protein transferases (FPTase and two geranylgeranyl-protein transferases), amino acid changes homologous to those found in mutant yeast prenyl-protein transferase subunits were made in the subunits of human FPTase. Substitutions in either the alpha or beta subunits that decrease the activity of yeast prenyl-protein transferases were also observed to impair human FPTase. Kinetic analyses showed that these mutant human FPTases have Km and kcat values that are altered with respect to wild-type human FPTase.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | *Alkyl and Aryl Transferases Amino Acid Sequence Animals Base Sequence Cattle *Cloning, Molecular DNA/chemistry Escherichia coli/genetics Farnesyltranstransferase *Gene Expression Humans Kinetics Molecular Sequence Data Mutation Polyisoprenyl Phosphates/*metabolism Rats Recombinant Proteins/chemistry/metabolism Saccharomyces cerevisiae/*enzymology Sequence Homology, Amino Acid Sesquiterpenes Structure-Activity Relationship Transferases/chemistry/*genetics/metabolism |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs > Powers lab |
| Depositing User: | Matt Covey |
| Date: | 18 May 1993 |
| Date Deposited: | 25 Feb 2014 21:29 |
| Last Modified: | 25 Feb 2014 21:29 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29527 |
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