A growth hormone agonist produced by targeted mutagenesis at binding site 1. Evidence that site 1 regulates bioactivity

J Biol Chem. 1995 Jul 14;270(28):16833-9. doi: 10.1074/jbc.270.28.16833.

Abstract

Growth hormone (GH) is believed to signal by dimerizing its receptor through two binding sites on the hormone. Previous attempts to increase the biopotency of GH by increasing its site 1 affinity have been unsuccessful, which has led to a bias toward engineering site 2 interactions in the quest for creation of super agonists. Here we report that increasing site 1 affinity can markedly increase proliferative bioactivity in FDC-P1 cells expressing full-length GHR. In contrast, we find three site 1 mutants with affinities for site one similar to or greater than wild type GH, which have markedly decreased bioactivity. Through crystal structure analysis of the receptor interactive regions of these GH analogues, we are able to suggest why previous mutagenesis on human GH failed to improve biopotency, and thus provide a new avenue for GH and cytokine agonist design.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cell Line
  • Growth Hormone / analogs & derivatives*
  • Growth Hormone / chemistry
  • Growth Hormone / metabolism
  • Humans
  • Mutagenesis, Site-Directed
  • Rabbits
  • Receptors, Somatotropin / metabolism
  • Structure-Activity Relationship
  • Thymidine / metabolism
  • Transfection

Substances

  • Receptors, Somatotropin
  • Growth Hormone
  • Thymidine