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. 2010 Nov 5;12(21):4848-51.
doi: 10.1021/ol101981b.

Efficient microwave-assisted synthesis of human islet amyloid polypeptide designed to facilitate the specific incorporation of labeled amino acids

Peter Marek  1 , Ann Marie Woys, Kelvin Sutton, Martin T Zanni, Daniel P Raleigh
Affiliations

Efficient microwave-assisted synthesis of human islet amyloid polypeptide designed to facilitate the specific incorporation of labeled amino acids

Peter Marek et al. Org Lett. .

Abstract

A cost-efficient, time-reducing solid-phase synthesis of the amyloidogenic, 37 residue islet amyloid polypeptide (IAPP) is developed using two pseudoprolines (highlighted blue in sequence) in combination with microwave technology. A yield twice that obtained with conventional syntheses is realized. The utility of this protocol is demonstrated by the synthesis of a (13)C(18)O-labeled Ser-20 IAPP variant, a prohibitively expensive and chemically challenging site to label via other protocols. TEM analysis shows the peptide forms normal amyloid (abstract image).

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Figures

Figure 1
Figure 1
Primary sequence of IAPP, illustrating the coupling scheme used for the microwave solid-phase peptide synthesis. Residues that were double coupled are underlined. The pseudoproline residues are colored blue, and all other double coupled residues are underlined.
Figure 2
Figure 2
(a) Reversed-phase analytical HPLC trace and (b) MALDI spectrum of crude, reduced IAPP. The HPLC was run on a C18 Vydac analytical column at a flow rate of 1 mL/min with a gradient of 0–90% buffer B (80% acetonitrile, 0.045% HCl) over 90 min.
Figure 3
Figure 3
(a) Reversed-phase analytical HPLC trace and (b) MALDI mass spectrum of pure, oxidized IAPP. The HPLC was run on a C18 Vydac analytical column at a gradient of 0–90% buffer-B (80% acetonitrile, 0.045% HCl) over 90 min. The pure peptide elutes at 48 min, corresponding to 48% buffer-B.
Figure 4
Figure 4
Thioflavin-T monitored kinetics of IAPP synthesized by microwave (red) and conventional (black) synthesis.
See this image and copyright information in PMC

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