Chemical Synthesis of K33 Acetylated SUMO Protein†

doi:10.7503/cjcu20160417

Abstract

Abstract:

Sumoylated protein was found to reduce the protein binding force with the SUMO-interaction motif(SIM) of effector protein when K33 of SUMO was acetylated. To further study the structures and mechanism, a large amount of uniformity acetylated SUMO protein was greatly needed. In this work, SUMO protein and K33 acetylated SUMO protein was efficiently obtained using high temperature assisted solid-phase peptide synthesis(SPPS) technology combined with peptide hydrazide ligation. SUMO and K33 acetylated SUMO was identified with corrected molecular weight by the SDS-PAGE. It was also confirmed that the chemically synthe-tic protein and biologically expressed SUMO2 own similar homogeneity and secondary structure by the CD analysis. These results provided a foundation for the follow-up study of K33 acetylated SUMO protein.

Key words: Small ubquitin-related modifier, Acetylation, High temperature assisted solid-phase peptide synthesis, Peptide hydrazide, Native chemical ligation

CLC Number:

O629

TrendMD:

WANG Yehai, KONG Yifu, CHEN Chenchen, LI Yiming. Chemical Synthesis of K33 Acetylated SUMO Protein^†[J]. Chem. J. Chinese Universities, 2016, 37(11): 1987.

Figures/Tables 6

Scheme 1 Amino acid sequence of SUMO2(A) and the synthesis strategy of SUMO2 and K33-Ac-SUMO2(B)

Fig.1 HPLC trace(A, B, C) and MS(D, E, F) of SUMO2-1(A, D), K33-Ac-SUMO2-1(B, E) and SUMO2-2(C, F)

Fig.2 Ligation trace of SUMO2-1(A) and SUMO2-2(B), and MS(C) of SUMO2

Fig.3 Ligation trace of K33-Ac-SUMO2-1(A) and SUMO2-2(B), and MS(C) of K33-Ac-SUMO2

Fig.4 SDS-PAGE of expressed SUMO2(1), synthesized SUMO2(2) and synthesized K33-Ac-SUMO2(3)

Fig.5 CD spectra of expressed SUMO2, synthesized SUMO2 and synthesized K33-Ac-SUMO2

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