Solution structure of proinsulin

Connecting domain flexibility and prohormone processing

Yanwu Yang, Qing Xin Hua, Jin Liu, Eri H. Shimizu, Meredith H. Choquette, Robert Mackin, Michael A. Weiss

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The folding of proinsulin, the single-chain precursor of insulin, ensures native disulfide pairing in pancreatic β-cells. Mutations that impair folding cause neonatal diabetes mellitus. Although the classical structure of insulin is well established, proinsulin is refractory to crystallization. Here, we employ heteronuclear NMR spectroscopy to characterize a monomeric analogue. Proinsulin contains a native-like insulin moiety (A- and B-domains); the tethered connecting (C) domain (as probed by {1H}-15N nuclear Overhauser enhancements) is progressively less ordered. Although the BC junction is flexible, residues near the CA junction exhibit α-helical-like features. Relative to canonical α-helices, however, segmental 13Cα/β chemical shifts are attenuated, suggesting that this junction and contiguous A-chain residues are molten. We propose that flexibility at each C-domain junction facilitates prohormone processing. Studies of protease SPC3 (PC1/3) suggest that C-domain sequences contribute to cleavage site selection. The structure of proinsulin provides a foundation for studies of insulin biosynthesis and its impairment in monogenic forms of diabetes mellitus.

Original languageEnglish
Pages (from-to)7847-7851
Number of pages5
JournalJournal of Biological Chemistry
Volume285
Issue number11
DOIs
StatePublished - Mar 12 2010

Fingerprint

Proinsulin
Insulin
Medical problems
Processing
Diabetes Mellitus
Biomolecular Nuclear Magnetic Resonance
Site selection
Biosynthesis
Chemical shift
Crystallization
Disulfides
Refractory materials
Nuclear magnetic resonance spectroscopy
Molten materials
Peptide Hydrolases
Magnetic Resonance Spectroscopy
Mutation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Solution structure of proinsulin : Connecting domain flexibility and prohormone processing. / Yang, Yanwu; Hua, Qing Xin; Liu, Jin; Shimizu, Eri H.; Choquette, Meredith H.; Mackin, Robert; Weiss, Michael A.

In: Journal of Biological Chemistry, Vol. 285, No. 11, 12.03.2010, p. 7847-7851.

Research output: Contribution to journalArticle

Yang, Yanwu ; Hua, Qing Xin ; Liu, Jin ; Shimizu, Eri H. ; Choquette, Meredith H. ; Mackin, Robert ; Weiss, Michael A. / Solution structure of proinsulin : Connecting domain flexibility and prohormone processing. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 11. pp. 7847-7851.
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