Suppression of insulin aggregation by heparin.
The aggregation of insulin near its isoelectric point and at low ionic strength was suppressed in the presence of heparin. To understand this effect, we used turbidimetry and stopped-flow to study the pH- and ionic strength ( I)-dependence of the aggregation of heparin-free insulin. The results supported the role of interprotein electrostatic interactions, contrary to the commonly held view that such forces are minimized at pH = pI. Electrostatic modeling of insulin (DelPhi) revealed that attractive interactions arise from the marked charge anisotropy of insulin near pI. We show how screening of the interprotein attractions by added salt lead to maximum aggregation near I = 0.01 M, corresponding to a Debye length nearly equal to the diameter of the insulin dimer, consistent with a dipole-like protein charge distribution. This analysis is also consistent with suppression of aggregation by heparin, a strong polyanion that by binding to the positive domain of one protein, inhibits its interaction with the negative domain of another.
Giger
Katie
K
Department of Chemistry, Indiana University-Purdue University at Indianapolis, 402 North Blackford Street, Indianapolis, Indiana 46202, USA.
Vanam
Ram P
RP
Seyrek
Emek
E
Dubin
Paul L
PL
D000818:Animals; D003198:Computer Simulation; D006493:Heparin / Q000737:chemistry* / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000737:chemistry*; D007525:Isoelectric Focusing; D008958:Models, Molecular; D009391:Nephelometry and Turbidimetry; D009994:Osmolar Concentration; D011485:Protein Binding / Q000187:drug effects; D055672:Static Electricity; D013552:Swine
D007328:Insulin; D006493:Heparin