Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

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 Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1208668
Journal Information:
Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 50 Vol. 289; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

References (87)

Semisynthesis and properties of some insulin analogs journal September 1981
Physicochemical basis for the rapid time-action of Lys B28 Pro B29 -insulin: Dissociation of a protein-ligand complex journal December 1996
Mapping the Functional Surface of Insulin by Design: Structure and Function of a Novel A-Chain Analogue journal November 1996
A structural switch in a mutant insulin exposes key residues for receptor binding 1 1Edited by A. R. Fersht journal May 1998
Insulin Structure and Stability book January 1993
Galenics of Insulin book January 1987
Design of Non-Standard Insulin Analogs for the Treatment of Diabetes Mellitus journal September 2012
Amino acid sequence from degu islet amyloid-derived insulin shows unique sequence characteristics journal June 1990
Primary structure of human insulin-like growth factor II journal May 1978
The amino acid sequence of human insulin-like growth factor I and its structural homology with proinsulin. journal April 1978
Studies on the molecular organization of rat insulin secretory granules. journal December 1987
Importance of the character and configuration of residues B24, B25, and B26 in insulin-receptor interactions journal January 1991
Role of the Phenylalanine B24 Side Chain in Directing Insulin Interaction with Its Receptor journal April 1989
Spectroscopic signatures of the T to R conformational transition in the insulin hexamer journal November 1989
Cross-linking of a B25 azidophenylalanine insulin derivative to the carboxyl-terminal region of the alpha-subunit of the insulin receptor. Identification of a new insulin-binding domain in the insulin receptor. journal November 1994
Solution structure of human insulin-like growthfactor II. Relationship to receptor and binding protein interactions journal January 1995
Evolution of the insulin molecule: insights into structure-activity and phylogenetic relationships journal July 2001
Diabetes mellitus due to the toxic misfolding of proinsulin variants journal May 2013
Importance of main-chain flexibility and the insulin fold in insulin-receptor interactions journal July 1993
Mutations at the dimer, hexamer, and receptor-binding surfaces of insulin independently affect insulin-insulin and insulin-receptor interactions journal February 1992
On the role of methionine residues in the sequence-independent recognition of nonpolar protein surfaces journal July 1991
Disposition of the phenylalanine B25 side chain during insulin-receptor and insulin-insulin interactions journal August 1991
Conformational dynamics of insulin in solution. Circular dichroic studies journal October 1980
Probing the Mechanism of Insulin Fibril Formation with Insulin Mutants † journal July 2001
Crystal Structure of Human Insulin-like Growth Factor-1:  Detergent Binding Inhibits Binding Protein Interactions ‡ journal September 2001
Packing Is a Key Selection Factor in the Evolution of Protein Hydrophobic Cores † journal December 2001
Structural Origins of the Functional Divergence of Human Insulin-Like Growth Factor-I and Insulin † , ‡ journal July 2002
CHARMM Additive All-Atom Force Field for Carbohydrate Derivatives and Its Utility in Polysaccharide and Carbohydrate–Protein Modeling journal August 2011
P-LINCS:  A Parallel Linear Constraint Solver for Molecular Simulation journal December 2007
GROMACS 4:  Algorithms for Highly Efficient, Load-Balanced, and Scalable Molecular Simulation journal February 2008
Synthesis of destetrapeptide B27-30 human (porcine) insulin. Biologically active insulin analog journal September 1973
Studies of the Nucleation and Growth Reactions of Selected Types of Insulin Fibrils journal June 1953
All-Atom Empirical Potential for Molecular Modeling and Dynamics Studies of Proteins † journal April 1998
Toward Understanding Insulin Fibrillation journal May 1997
Assembly and Dissociation of Human Insulin and LysB28ProB29-Insulin Hexamers: A Comparison Study journal January 1997
Structure of Rhombohedral 2 Zinc Insulin Crystals journal November 1969
Surface and inside volumes in globular proteins journal February 1979
Monomeric insulins obtained by protein engineering and their medical implications journal June 1988
Phenol stabilizes more helix in a new symmetrical zinc insulin hexamer journal April 1989
Receptor binding redefined by a structural switch in a mutant human insulin journal November 1991
Structure of the insulin receptor ectodomain reveals a folded-over conformation journal September 2006
How insulin engages its primary binding site on the insulin receptor journal January 2013
Structural biology of insulin and IGF1 receptors: implications for drug design journal October 2002
Mutational analysis of the N-linked glycosylation sites of the human insulin receptor journal May 2000
Mutational analysis of the N-linked glycosylation sites of the human insulin receptor journal April 2000
Insulin Analogues journal January 2005
Canonical sampling through velocity rescaling journal January 2007
Molecular dynamics with coupling to an external bath journal October 1984
A smooth particle mesh Ewald method journal November 1995
The first three domains of the insulin receptor differ structurally from the insulin-like growth factor 1 receptor in the regions governing ligand specificity journal August 2006
Structural resolution of a tandem hormone-binding element in the insulin receptor and its implications for design of peptide agonists journal March 2010
Protective hinge in insulin opens to enable its receptor engagement journal August 2014
Semisynthesis and biological activity of porcine [LeuB24]insulin and [LeuB25]insulin. journal June 1980
Identification of a mutant human insulin predicted to contain a serine-for-phenylalanine substitution. journal December 1983
An Achilles' Heel in an Amyloidogenic Protein and Its Repair: INSULIN FIBRILLATION AND THERAPEUTIC DESIGN journal January 2010
Structural Integrity of the B24 Site in Human Insulin Is Important for Hormone Functionality journal February 2013
Biophysical Optimization of a Therapeutic Protein by Nonstandard Mutagenesis: STUDIES OF AN IODO-INSULIN DERIVATIVE journal July 2014
Characterization of the Functional Insulin Binding Epitopes of the Full-length Insulin Receptor journal March 2005
Proinsulin Is Refractory to Protein Fibrillation: TOPOLOGICAL PROTECTION OF A PRECURSOR PROTEIN FROM CROSS- β ASSEMBLY journal October 2005
Structural Basis for the Lower Affinity of the Insulin-like Growth Factors for the Insulin Receptor journal November 2007
Enhancing the Activity of a Protein by Stereospecific Unfolding: CONFORMATIONAL LIFE CYCLE OF INSULIN AND ITS EVOLUTIONARY ORIGINS journal March 2009
Decoding the Cryptic Active Conformation of a Protein by Synthetic Photoscanning: INSULIN INSERTS A DETACHABLE ARM BETWEEN RECEPTOR DOMAINS journal March 2009
Isolation and Properties of Secretory Granules from rat Islets of Langerhans journal April 1969
PROVAT: a tool for Voronoi tessellation analysis of protein structures and complexes journal June 2005
Voroprot: an interactive tool for the analysis and visualization of complex geometric features of protein structure journal December 2010
The structure of 2Zn pig insulin crystals at 1.5 Å resolution
  • Baker, Edward N.; Blundell, Thomas Leon; Cutfield, John F.
  • Philosophical Transactions of the Royal Society of London. B, Biological Sciences, Vol. 319, Issue 1195, p. 369-456 https://doi.org/10.1098/rstb.1988.0058
journal July 1988
R 6 hexameric insulin complexed with m -cresol or resorcinol journal December 2000
Integration, scaling, space-group assignment and post-refinement journal January 2010
PHENIX: a comprehensive Python-based system for macromolecular structure solution journal January 2010
Solvation and the hidden thermodynamics of a zinc finger probed by nonstandard repair of a protein crevice journal December 2004
Enzymatic semisynthesis of porcine despentapeptide (B26-30) insulin using unprotected desoctapeptide (B23-30) insulin as a substrate: Model studies journal May 1986
Three-dimensional structure of human insulin-like growth factor-I (IGF-I) determined by 1H-NMR and distance geometry journal May 1993
Continuous glucose monitoring and closed-loop systems journal January 2006
Editorial journal July 2006
Atomic-Level Characterization of the Structural Dynamics of Proteins journal October 2010
Linking Crystallographic Model and Data Quality journal May 2012
Response of a protein structure to cavity-creating mutations and its relation to the hydrophobic effect journal January 1992
Deciphering the message in protein sequences: tolerance to amino acid substitutions journal March 1990
Aromatic-aromatic interaction: a mechanism of protein structure stabilization journal July 1985
Structure and Evolution of the Insulin Gene journal December 1985
Human insulin B24 (Phe----Ser). Secretion and metabolic clearance of the abnormal insulin in man and in a dog model. journal May 1984
Cloning of Complementary DNAs Encoding Islet Amyloid Polypeptide, Insulin, and Glucagon Precursors from a New World Rodent, the Degu, Octodon degus journal August 1990
Mutant INS-Gene Induced Diabetes of Youth: Proinsulin Cysteine Residues Impose Dominant-Negative Inhibition on Wild-Type Proinsulin Transport journal October 2010
Structure and Activity of Insulin, XVl. Semisyntheses of Desheptapeptide-(B24—30)- up to Destripeptide-(B28—30)-Insulin with Lysine or Alanine in Place of Arginine in Position B22: Influence on the Three-Step-Increase of Activity in Positions B24—26 (Phe-Phe-Tyr) journal January 1978
Insulin Self-Association and the Relationship to Pharmacokinetics and Pharmacodynamics journal January 2001
CHANGES IN RECEPTOR BINDING, BIOLOGICAL ACTIVITY AND IMMUNOREACTIVITY OF INSULIN CAUSED BY REPLACING THE RESIDUES B23-B26 VVITH ALANINE journal January 1984
Decreased Biologic Activity and Degradation of Human [SerB24]-Insulin, a Second Mutant Insulin journal June 1985

Cited By (5)

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 journal April 2016
A thing of beauty: Structure and function of insulin's “aromatic triplet” journal September 2018
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 text January 2016

Similar Records

Importance of the character and configuration of residues B24, B25, and B26 in insulin-receptor interactions
Journal Article · Thu Jan 24 23:00:00 EST 1991 · Journal of Biological Chemistry; (USA) · OSTI ID:6057913
Mutations at the dimer, hexamer, and receptor-binding surfaces of insulin independently affect insulin-insulin and insulin-receptor interactions
Journal Article · Mon Feb 17 23:00:00 EST 1992 · Biochemistry; (United States) · OSTI ID:5559639
Efficient Total Chemical Synthesis of 13C= 18O Isotopomers of Human Insulin for Isotope-Edited FTIR
Journal Article · Tue Dec 29 19:00:00 EST 2015 · ChemBioChem: a European journal of chemical biology · OSTI ID:1241077

Related Subjects

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