Analysis of ligand-receptor cross-linked fragments by mass spectrometry

2005-03-01
Son, Çağdaş Devrim
Hurst, GB
Naider, F
Becker, JM
G-protein coupled receptors (GPCRs) are a class of integral membrane receptor proteins that are characterized by a signature seven-transmembrane (7-TM) configuration. The alpha-factor receptor (Ste2p) from Saccharomyces cerevisiae is a GPCR that, upon binding of a peptide ligand, transduces a signal to initiate a cascade of events leading to the mating of haploid yeast cells. This study summarizes the application of affinity purification and of matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) experiments using biotinylated photoactivatable alpha-factor analogs. Affinity purification and enrichment of biotinylated peptides by monomeric avidin beads resulted in mass spectrometric detection of specific signals corresponding to crosslinked fragments of Ste2p. Data obtained from cyanogen bromide (CNBr) fragments of receptor cross-linked to an alpha-factor analog with the photoaffinity group p-benzoyl-L-phenylalanine on position 1 were in agreement with the previous results reported by our laboratory suggesting the cross-linking between position 1 of alpha-factor and a region of Ste2p covering residues 251-294.
JOURNAL OF PEPTIDE RESEARCH

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Citation Formats
Ç. D. Son, G. Hurst, F. Naider, and J. Becker, “Analysis of ligand-receptor cross-linked fragments by mass spectrometry,” JOURNAL OF PEPTIDE RESEARCH, pp. 418–426, 2005, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/40891.