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Substrate specificity determinants in the farnesyltransferase beta-subunit

C E Trueblood1, V L Boyartchuk, J Rine

  • 1Division of Genetics, Department of Molecular and Cell Biology, 401 Barker Hall, University of California, Berkeley, CA 94720, USA.

Proceedings of the National Academy of Sciences of the United States of America
|October 6, 1997
PubMed
Summary

Yeast farnesyltransferase specificity was investigated. Oppositely charged amino acids at key positions enhance protein prenylation, suggesting electrostatic interactions guide substrate binding.

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Area of Science:

  • Molecular Biology
  • Enzymology
  • Protein Modification

Background:

  • Protein prenyltransferases attach isoprenoid lipids to proteins.
  • This modification is crucial for protein function and localization.
  • Understanding enzyme-substrate specificity is key to protein prenylation research.

Purpose of the Study:

  • To explore specificity determinants in Saccharomyces cerevisiae farnesyltransferase.
  • To investigate interactions between the enzyme's beta-subunit and CaaX protein substrates.

Main Methods:

  • Systematic substitutions at amino acid 149 of the farnesyltransferase beta-subunit.
  • Substitutions were made at the C-terminal amino acid of CaaX protein substrates (Ras2p and a-factor).
  • Assessed prenylation efficiency based on these combined substitutions.

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Main Results:

  • Efficient prenylation occurred when amino acid 149 and the substrate's C-terminal amino acid had opposite charges.
  • Prenylation was inefficient when these positions had like charges.
  • Demonstrated a clear electrostatic interaction governing enzyme-substrate binding.

Conclusions:

  • Electrostatic interactions between yeast farnesyltransferase beta-subunit (amino acid 149) and CaaX protein substrates are critical for specificity.
  • The C-terminal amino acid of the protein substrate likely binds near amino acid 149 of the enzyme.
  • Provides insights into the molecular mechanisms of protein prenylation.