Glycosylation of the enhanced aromatic sequon is similarly stabilizing in three distinct reverse turn contexts

被引:60
作者
Price, Joshua L. [1 ,2 ]
Powers, David L. [3 ]
Powers, Evan T. [1 ,2 ]
Kelly, Jeffery W. [1 ,2 ,4 ]
机构
[1] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
[2] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
[3] Clarkson Univ, Dept Math, Potsdam, NY 13699 USA
[4] Scripps Res Inst, Dept Mol & Expt Med, La Jolla, CA 92037 USA
基金
美国国家卫生研究院;
关键词
glycoprotein; biopharmaceutical; conjugate; native state stabilization; beta-sheet; ASPARAGINE-LINKED GLYCOSYLATION; RECOMBINANT-HUMAN-ERYTHROPOIETIN; THERAPEUTIC PROTEINS; BETA-SHEET; WW DOMAIN; PEPTIDE CONFORMATION; N-GLYCOSYLATION; HALF-LIFE; GLYCAN; CARBOHYDRATE;
D O I
10.1073/pnas.1105880108
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Cotranslational N-glycosylation can accelerate protein folding, slow protein unfolding, and increase protein stability, but the molecular basis for these energetic effects is incompletely understood. N-glycosylation of proteins at naive sites could be a useful strategy for stabilizing proteins in therapeutic and research applications, but without engineering guidelines, often results in unpredictable changes to protein energetics. We recently introduced the enhanced aromatic sequon as a family of portable structural motifs that are stabilized upon glycosylation in specific reverse turn contexts: a five-residue type I beta-turn harboring a G1 beta-bulge (using a Phe-Yyy-Asn-Xxx-Thr sequon) and a type II beta-turn within a six-residue loop (using a Phe-Yyy-Zzz-Asn-Xxx-Thr sequon) [Culyba EK, et al. (2011) Science 331: 571-575]. Here we show that glycosylating a new enhanced aromatic sequon, Phe-Asn-Xxx-Thr, in a type I' beta-turn stabilizes the Pin 1 WW domain. Comparing the energetic effects of glycosylating these three enhanced aromatic sequons in the same host WW domain revealed that the glycosylation-mediated stabilization is greatest for the enhanced aromatic sequon complementary to the type I beta-turn with a G1 beta-bulge. However, the portion of the stabilization from the tripartite interaction between Phe, Asn(GlcNAc), and Thr is similar for each enhanced aromatic sequon in its respective reverse turn context. Adding the Phe-Asn-Xxx-Thr motif (in a type I' beta-turn) to the enhanced aromatic sequon family doubles the number of proteins that can be stabilized by glycosylation without having to alter the native reverse turn type.
引用
收藏
页码:14127 / 14132
页数:6
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