Separation and Identification of Oyster Peptide Modified by Plastein Reaction and Characterization of Peptide-zinc Complexes†

doi:10.7503/cjcu20170570

Abstract

Abstract:

An oyster peptide modified by plastein reaction was isolated from the modified product using different methods including Sephadex G-15 gelcolumn chromatography and reversed-phase high-performance liquid chromalography(RP-HPLC). The purified peptide was sequenced as EVPPEEH with a zinc chelate activity of 161 mg/g. The peptide sequence was used as template to synthesize the pure peptide which was used to prepare peptide-zinc complexes. The results of infrared spectroscopy and circular dichroism of the complexes showed that the carbonyl oxygen belonging to the peptide chain was the primary binding site for Zn²⁺. Compared with the pure peptide, the irregular curl and β fold of complexes was reduced while β angle increases. Molecular mechanics simulation experiment and secondary spectrum of complexes showed that there were two kinds of spatial conformation of complexes. One way was to chelate a zinc ion by six coordination where the main chelating site was the carbonyl oxygen between Val-2 and Pro-3 or Clu-5 and Clu-6. The other was to chelate a zinc ion by four coordination where the main chelating site was the carbonyl oxygen between Clu-5 and Clu-6.

Key words: Plastein reaction, Chelate activity, Isolation and purification, Chelate site

CLC Number:

O629.72

TrendMD:

CAO Yuhui, ZHANG Juanjuan, WANG Zaiyang, ZHAO Yuanhui. Separation and Identification of Oyster Peptide Modified by Plastein Reaction and Characterization of Peptide-zinc Complexes^†[J]. Chem. J. Chinese Universities, 2018, 39(3): 470.

Figures/Tables 7

Fig.1 Sephadex G-15 chromatograph of phenylalanine standard(A) and oyster peptides(B)

Fig.2 Isolation and purification of modified oyster peptides by plastein reaction and zinc ion chelating activity of different components(A) a—d: Modified peptides isolated by Sephadex G-15 chromatograph; (B) zinc ion chelating activity of components a—d; (C) further purified components a1 and a2 by RP-HPLC on Zorbax SB-C18 column; (D) zinc ion chelating activity of components a1 and a2.

Fig.3 Identification of the amino acid sequence of component a1 using LC/LTQ mass spectrometry

Fig.4 FTIR spectra of the pure peptide(EVPPEEH)(a) and EVPPEEH-zinc complex(b)

Fig.5 CD spectra of the pure peptide(EVPPEEH)(a) and EVPPEEH-zinc complex(b)

Fig.6 Molecular mechanics simulation of EVPPEEH-zinc complex

Fig.7 Mass spectra of EVPPEEH(A), EVPPEEH-Zn(B) and MS/MS spectrum of EVPPEEH-Zn(C)

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