Identification of a Contact Site for Residue 19 of Parathyroid Hormone (PTH) and PTH-Related Protein Analogs in Transmembrane Domain Two of the Type 1 PTH Receptor

Robert C. Gensure, Naoto Shimizu, Janet Tsang, Thomas J. Gardella

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Recent functional studies have suggested that position 19 in PTH interacts with the portion of the PTH-1 receptor (P1R) that contains the extracellular loops and seven transmembrance helices (TMs) (the J domain). We tested this hypothesis using the photoaffinity cross-linking approach. A PTHrP(1-36) analog and a conformationally constrained PTH(1-21) analog, each containing para-benzoyl-L-phenylalanine (Bpa) at position 19, each cross-linked efficiently to the P1R expressed in COS-7 cells, and digestive mapping analysis localized the cross-linked site to the interval (Leu232-Lys240) at the extracellular end of TM2. Point mutation analysis identified Ala234, Val235, and Lys240 as determinants of cross-linking efficiency, and the Lys240→Ala mutation selectively impaired the binding of PTH(1-21) and PTH(1-19) analogs, relative to that of PTH(1-15) analogs. The findings support the hypothesis that residue 19 of the receptor-bound ligand contacts, or is close to, the P1R J domain-specifically, Lys240 at the extracellular end of TM2. The findings also support a molecular model in which the 1-21 region of PTH binds to the extracellular face of the P1R J domain as an α-helix.

Original languageEnglish (US)
Pages (from-to)2647-2658
Number of pages12
JournalMolecular Endocrinology
Volume17
Issue number12
DOIs
StatePublished - Dec 2003
Externally publishedYes

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Parathyroid Hormone Receptor Type 1
Parathyroid Hormone-Related Protein
Parathyroid Hormone
Molecular Models
COS Cells
Phenylalanine
Point Mutation
Ligands
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Identification of a Contact Site for Residue 19 of Parathyroid Hormone (PTH) and PTH-Related Protein Analogs in Transmembrane Domain Two of the Type 1 PTH Receptor. / Gensure, Robert C.; Shimizu, Naoto; Tsang, Janet; Gardella, Thomas J.

In: Molecular Endocrinology, Vol. 17, No. 12, 12.2003, p. 2647-2658.

Research output: Contribution to journalArticle

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