The C-terminus of the B-chain of human insulin-like peptide 5 is critical for cognate RXFP4 receptor activity

Nitin A. Patil, Ross A D Bathgate, Martina Kocan, Sheng Yu Ang, Julien Tailhades, Frances Separovic, Roger Summers, Johannes Grosse, Richard A. Hughes, John D. Wade, Mohammed Akhter Hossain

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Insulin-like peptide 5 (INSL5) is an orexigenic peptide hormone belonging to the relaxin family of peptides. It is expressed primarily in the L-cells of the colon and has a postulated key role in regulating food intake. Its G protein-coupled receptor, RXFP4, is a potential drug target for treating obesity and anorexia. We studied the effect of modification of the C-terminus of the A and B-chains of human INSL5 on RXFP4 binding and activation. Three variants of human INSL5 were prepared using solid phase peptide synthesis and subsequent sequential regioselective disulfide bond formation. The peptides were synthesized as C-terminal acids (both A- and B-chains with free C-termini, i.e., the native form), amides (both chains as the C-terminal amide) and one analog with the C-terminus of its A-chain as the amide and the C-terminus of the B-chain as the acid. The results showed that C-terminus of the B-chain is more important than that of the A-chain for RXFP4 binding and activity. Amidation of the A-chain C-terminus does not have any effect on the INSL5 activity. The difference in RXFP4 binding and activation between the three peptides is believed to be due to electrostatic interaction of the free carboxylate of INSL5 with a positively charged residue (s), either situated within the INSL5 molecule itself or in the receptor extracellular loops.

Original languageEnglish
Pages (from-to)987-992
Number of pages6
JournalAmino Acids
Volume48
Issue number4
DOIs
Publication statusPublished - 1 Apr 2016

Keywords

  • INSL5
  • Insulin-like peptide 5
  • Peptide
  • Relaxin

Cite this

Patil, Nitin A. ; Bathgate, Ross A D ; Kocan, Martina ; Ang, Sheng Yu ; Tailhades, Julien ; Separovic, Frances ; Summers, Roger ; Grosse, Johannes ; Hughes, Richard A. ; Wade, John D. ; Hossain, Mohammed Akhter. / The C-terminus of the B-chain of human insulin-like peptide 5 is critical for cognate RXFP4 receptor activity. In: Amino Acids. 2016 ; Vol. 48, No. 4. pp. 987-992.
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abstract = "Insulin-like peptide 5 (INSL5) is an orexigenic peptide hormone belonging to the relaxin family of peptides. It is expressed primarily in the L-cells of the colon and has a postulated key role in regulating food intake. Its G protein-coupled receptor, RXFP4, is a potential drug target for treating obesity and anorexia. We studied the effect of modification of the C-terminus of the A and B-chains of human INSL5 on RXFP4 binding and activation. Three variants of human INSL5 were prepared using solid phase peptide synthesis and subsequent sequential regioselective disulfide bond formation. The peptides were synthesized as C-terminal acids (both A- and B-chains with free C-termini, i.e., the native form), amides (both chains as the C-terminal amide) and one analog with the C-terminus of its A-chain as the amide and the C-terminus of the B-chain as the acid. The results showed that C-terminus of the B-chain is more important than that of the A-chain for RXFP4 binding and activity. Amidation of the A-chain C-terminus does not have any effect on the INSL5 activity. The difference in RXFP4 binding and activation between the three peptides is believed to be due to electrostatic interaction of the free carboxylate of INSL5 with a positively charged residue (s), either situated within the INSL5 molecule itself or in the receptor extracellular loops.",
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The C-terminus of the B-chain of human insulin-like peptide 5 is critical for cognate RXFP4 receptor activity. / Patil, Nitin A.; Bathgate, Ross A D; Kocan, Martina; Ang, Sheng Yu; Tailhades, Julien; Separovic, Frances; Summers, Roger; Grosse, Johannes; Hughes, Richard A.; Wade, John D.; Hossain, Mohammed Akhter.

In: Amino Acids, Vol. 48, No. 4, 01.04.2016, p. 987-992.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - The C-terminus of the B-chain of human insulin-like peptide 5 is critical for cognate RXFP4 receptor activity

AU - Patil, Nitin A.

AU - Bathgate, Ross A D

AU - Kocan, Martina

AU - Ang, Sheng Yu

AU - Tailhades, Julien

AU - Separovic, Frances

AU - Summers, Roger

AU - Grosse, Johannes

AU - Hughes, Richard A.

AU - Wade, John D.

AU - Hossain, Mohammed Akhter

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AB - Insulin-like peptide 5 (INSL5) is an orexigenic peptide hormone belonging to the relaxin family of peptides. It is expressed primarily in the L-cells of the colon and has a postulated key role in regulating food intake. Its G protein-coupled receptor, RXFP4, is a potential drug target for treating obesity and anorexia. We studied the effect of modification of the C-terminus of the A and B-chains of human INSL5 on RXFP4 binding and activation. Three variants of human INSL5 were prepared using solid phase peptide synthesis and subsequent sequential regioselective disulfide bond formation. The peptides were synthesized as C-terminal acids (both A- and B-chains with free C-termini, i.e., the native form), amides (both chains as the C-terminal amide) and one analog with the C-terminus of its A-chain as the amide and the C-terminus of the B-chain as the acid. The results showed that C-terminus of the B-chain is more important than that of the A-chain for RXFP4 binding and activity. Amidation of the A-chain C-terminus does not have any effect on the INSL5 activity. The difference in RXFP4 binding and activation between the three peptides is believed to be due to electrostatic interaction of the free carboxylate of INSL5 with a positively charged residue (s), either situated within the INSL5 molecule itself or in the receptor extracellular loops.

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