A Photoactive Phosphonamide Derivative of GTP for the Identification of the GTP-Binding Domain in ݭTubulin

Ashok J. Chavan, Hyuntae Kim, Boyd E. Haley, David S. Watt

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

A GTP photoaffinity probe (125I-APTG) was developed that incorporated an [125I]-N-(4-azidophenyl)-2-amino-3-(4-hydroxy-3-iodophenyl)propionamide group at the ?-position of GTP through a phos-phonamide linkage. A combination of saturation and GTP protection studies (90% protection at 25 µM GTP with an apparent of 5 µM) validated the use of this new probe as a satisfactory GTP mimic. This probe offered the advantage of possessing an 125I radiolabel external to the GTP moiety, in contrast to the previously reported [?32P]-8-N3GTP that possessed an internal 32P radiolabel. This novel feature accommodated the purification of photolabeled peptides using a combination of ion-exclusion, gel filtration, and HPLC techniques. [125I]APTG was used to identify a peptide (ݺ65-79) in the exchangeable GTP-binding domain of the ݭsubunit of tubulin.

Original languageEnglish
Pages (from-to)337-344
Number of pages8
JournalBioconjugate Chemistry
Volume1
Issue number5
DOIs
StatePublished - Sep 1 1990

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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