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Title: Aromatic anchor at an invariant hormone-receptor interface: Function of insulin residue B24 with application to protein design

Journal Article · · Journal of Biological Chemistry
 [1];  [2];  [1];  [1];  [1];  [1];  [3];  [1];  [1];  [1];  [4];  [1]
  1. Case Western Reserve Univ., Cleveland, OH (United States)
  2. La Trobe Univ., Melbourne, VIC (Australia)
  3. Walter and Eliza Hall Institute of Medical Research, Parkville, VIC (Australia)
  4. Walter and Eliza Hall Institute of Medical Research, Parkville, VIC (Australia); Univ. of Melbourne, Parkville, VIC (Australia)
+ Show Author Affiliations

Crystallographic studies of insulin bound to fragments of the insulin receptor have recently defined the topography of the primary hormone-receptor interface. Here, we have investigated the role of PheB24, an invariant aromatic anchor at this interface and site of a human mutation causing diabetes mellitus. An extensive set of B24 substitutions has been constructed and tested for effects on receptor binding. Although aromaticity has long been considered a key requirement at this position, MetB24 was found to confer essentially native affinity and bioactivity. Molecular modeling suggests that this linear side chain can serve as an alternative hydrophobic anchor at the hormone-receptor interface. These findings motivated further substitution of PheB24 by cyclohexanylalanine (Cha), which contains a nonplanar aliphatic ring. Contrary to expectations, [ChaB24]insulin likewise exhibited high activity. Furthermore, its resistance to fibrillation and the rapid rate of hexamer disassembly, properties of potential therapeutic advantage, were enhanced. The crystal structure of the ChaB24 analog, determined as an R6 zinc-stabilized hexamer at a resolution of 1.5 Å, closely resembles that of wild-type insulin. The nonplanar aliphatic ring exhibits two chair conformations with partial occupancies, each recapitulating the role of PheB24 at the dimer interface. Together, these studies have defined structural requirements of an anchor residue within the B24-binding pocket of the insulin receptor; similar molecular principles are likely to pertain to insulin-related growth factors. Finally, our results highlight in particular the utility of nonaromatic side chains as probes of the B24 pocket and suggest that the nonstandard Cha side chain may have therapeutic utility.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1208668
Journal Information:
Journal of Biological Chemistry, Vol. 289, Issue 50; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 18 works
Citation information provided by
Web of Science

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Cited By (5)

Analysis of Insulin Analogs and the Strategy of Their Further Development journal January 2018
Diabetes mellitus caused by mutations in human insulin: analysis of impaired receptor binding of insulins Wakayama , Los Angeles and Chicago using pharmacoinformatics journal April 2016
A thing of beauty: Structure and function of insulin's “aromatic triplet” journal September 2018
Extending Halogen-based Medicinal Chemistry to Proteins: IODO-INSULIN AS A CASE STUDY journal November 2016
Diabetes mellitus caused by mutations in human insulin: analysis of impaired receptor binding of insulins Wakayama, Los Angeles and Chicago using pharmacoinformatics text January 2016

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
diabetes
mutagenesis
protein design
protein structure
receptor tyrosine kinase hormone
nonstandard mutagenesis