Photoaffinity labeling of the lutropin receptor with synthetic peptide for carboxyl terminus of the human choriogonadotropin α subunit

Gopal C. Kundu, Inhae Ji, Daniel J Mc Cormick, Tae H. Ji

Research output: Contribution to journalArticle

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Abstract

Human choriogonadotropin (hCG) consists of an α subunit and a β subunit. The existing evidence from various studies using truncation, substitution, synthetic hormone peptides, and hCG crystals suggests that the C-terminal region of the α subunit contacts the luteinizing hormone/choriogonoadotropin (LH/CG) receptor and is involved in receptor activation. Despite a deluge of the speculation and the important role of the αC-terminal region, direct evidence for its interaction with the receptor has been elusive. Because of the significant biological activity, it is imperative to prove the interaction of the αC-terminal region. For this purpose, decamer peptides corresponding to the α subunit sequence from His83 to Ser9283-92) were derivatized with the N-hydroxysuccinimide ester of 4- azidobenzoylglycine (ABG) and radioiodinated. The resulting ABG-125I- α83-92 was capable of binding and activating the LH/CG receptor. Furthermore, UV-sensitive ABG-125I-α83-92 exclusively photoaffinity- labeled an ~86-kDa molecule. This labeled molecule was shown to be the LH/CG receptor by various methods including immunoprecipitation by anti-LH/CG receptor antiserum. In addition, evidence is presented that the amino group of αLys91 of α83-92 is in such close proximity to a carboxyl group of the receptor that this pair is cross-linked to form an amide, a zero length cross-link. This low affinity contact of α83-92 and the receptor is sufficient for receptor activation and is crucial for the full understanding of the mechanistics of the receptor activation steps.

Original languageEnglish (US)
Pages (from-to)11063-11066
Number of pages4
JournalJournal of Biological Chemistry
Volume271
Issue number19
DOIs
StatePublished - 1996

Fingerprint

LH Receptors
Chorionic Gonadotropin
Labeling
Peptides
Chemical activation
Molecules
Peptide Hormones
Bioactivity
Immunoprecipitation
Amides
Immune Sera
Substitution reactions
Crystals

ASJC Scopus subject areas

  • Biochemistry

Cite this

Photoaffinity labeling of the lutropin receptor with synthetic peptide for carboxyl terminus of the human choriogonadotropin α subunit. / Kundu, Gopal C.; Ji, Inhae; Mc Cormick, Daniel J; Ji, Tae H.

In: Journal of Biological Chemistry, Vol. 271, No. 19, 1996, p. 11063-11066.

Research output: Contribution to journalArticle

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