P.F.S.R.L. - AMYLIN


Introduction

One of the major pathologies associated with non-insulin-dependent diabetes mellitus (NIDDM) is the formation of amyloid deposits, composed of high molecular weight aggregates of the islet amyloid polypeptide (IAPP). In our study we use an aggregable fragment of human IAPP (hIAPP) (amino acids 20-29) to investigate the effects of electrostatic interactions on the aggregation properties of the peptide, as well as the effects of trifluoroethanol (TFE) on peptide aggregation and conformation. We monitor the aggregation of the hIAPP fragment in solutions of pH ranging from 4.6 to 9.4. The aggregation rate peaks at pH of ca 5-6, indicating that the charged groups that are titrated at both high and low pH facilitate the aggregation. The aggregation of the hIAPP fragments has also been monitored in neutral buffer containing various concentrations of NaCl. Charge shielding by increasing NaCl concentrations resulted in prolonged lag time to aggregation. Fractional concentrations of TFE as low as 1% (v/v) in solutions were found to inhibit aggregation of the peptide completely. Circular dichroism (CD) studies of the hIAPP fragment in various concentrations of TFE show that structural shifts in the fragment occur very slowly, as long as seven days after addition of TFE to the peptide solutions.



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