Journal of Biomolecular NMR

, Volume 40, Issue 1, pp 55–64 | Cite as

Structure of human insulin monomer in water/acetonitrile solution

  • Wojciech Bocian
  • Jerzy Sitkowski
  • Elżbieta Bednarek
  • Anna Tarnowska
  • Robert Kawęcki
  • Lech Kozerski


Here we present evidence that in water/acetonitrile solvent detailed structural and dynamic information can be obtained for important proteins that are naturally present as oligomers under native conditions. An NMR-derived human insulin monomer structure in H2O/CD3CN, 65/35 vol%, pH 3.6 is presented and compared with the available X-ray structure of a monomer that forms part of a hexamer (Acta Crystallogr. 2003 Sec. D59, 474) and with NMR structures in water and organic cosolvent. Detailed analysis using PFGSE NMR, temperature-dependent NMR, dilution experiments and CSI proves that the structure is monomeric in the concentration and temperature ranges 0.1–3 mM and 10–30°C, respectively. The presence of long-range interstrand NOEs, as found in the crystal structure of the monomer, provides the evidence for conservation of the tertiary structure. Starting from structures calculated by the program CYANA, two different molecular dynamics simulated annealing refinement protocols were applied, either using the program AMBER in vacuum (AMBER_VC), or including a generalized Born solvent model (AMBER_GB).


Insulin monomer MD in explicit solvent NMR structure Water/acetonitrile solvent 



A. T. gratefully acknowledges the financial support during this work by means of Ministry of Science and Higher Education grant no. 0278/P01/2006/30.

Supplementary material

10858_2007_9206_MOESM1_ESM.pdf (4.9 mb)
(PDF 5042 kb)


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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Wojciech Bocian
    • 1
    • 2
  • Jerzy Sitkowski
    • 1
    • 2
  • Elżbieta Bednarek
    • 1
  • Anna Tarnowska
    • 2
  • Robert Kawęcki
    • 2
  • Lech Kozerski
    • 1
    • 2
  1. 1.National Medicines InstituteWarsawPoland
  2. 2.Institute of Organic Chemistry Polish Academy of SciencesWarsawPoland

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