Synthesis and characterization of novel biotinylated carboxyl-terminal parathyroid hormone peptides that specifically crosslink to the CPTH-receptor

Santanu Banerjee, Hafez Selim, Gihan Suliman, Andrew I. Geller, Harald Jüppner, F. Richard Bringhurst, Paola Divieti

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Parathyroid hormone (PTH) regulates calcium, phosphorous and skeletal homeostasis via interaction with the G protein-coupled PTH/PTHrP receptor, which is fully activated by the amino-terminal 34 amino-acid portion of the hormone. Recent evidence points to the existence of another class of receptors for PTH that recognize the carboxyl (C)-terminal region of intact PTH (1-84) (CPTHRs) and are highly expressed by osteocytes. Here we report the synthesis and characterization of two novel bifunctional CPTH ligands that include benzoylphenylalanine (Bpa) substitutions near their amino-termini and carboxyl-terminal biotin moieties, as well as a tyrosine34 substitution to enable radioiodination. These peptides are shown to bind to CPTHRs with affinity similar to that of PTH (1-84) and to be specifically and covalently crosslinked to CPTHRs upon exposure to ultraviolet light. Crosslinking to osteocytes or osteoblastic cells generates complexes of 80 and 220 kDa, of which the larger form represents an aggregate that can be resolved into the 80 kDa. The crosslinked products can be further purified using immunoaffinity and avidin-based affinity procedures. While the molecular structure of the CPTHR(s) remains undefined, these bifunctional ligands represent powerful new tools for use in isolating and characterizing CPTHR protein(s).

Original languageEnglish (US)
Pages (from-to)3352-3362
Number of pages11
JournalPeptides
Volume27
Issue number12
DOIs
StatePublished - Dec 2006

Bibliographical note

Funding Information:
The authors would like to thank Dr. Henry M Kronenberg for his critical suggestions during the course of this study. This work was supported by the National Institute of Health Grants DK-02889 and DK-65032 to P.D. and AR 47062 to F.R.B. and by an educational grant from NPS Pharmaceuticals.

Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.

Keywords

  • Benzoylphenylalanine
  • Carboxy-terminal parathyroid hormone
  • Parathyroid hormone
  • Parathyroid hormone receptor type-1

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