Matsushita Takahiko, Hinou Hiroshi, Fumoto Masataka, Kurogochi Masaki, Fujitani Naoki, Shimizu Hiroki, Nishimura Shin-ichiro
Division of Biological Sciences, Graduate School of Science, Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, 001-0021 Japan.
J Org Chem. 2006 Apr 14;71(8):3051-63. doi: 10.1021/jo0526643.
A MUC1-related glycopeptide having five core-2 hexasaccharide branches (C330H527N46O207, MW = 8450.9) was synthesized by a new strategy using a combination of microwave-assisted solid-phase synthesis (MA-SPGS) and enzymatic sugar elongation. Synthesis of a key glycopeptide intermediate was best achieved in a combination of PEGA [poly(ethylene glycol)-poly-(N,N-dimethylacrylamide) copolymer] resin and MA-SPGS using glycosylated amino acid building blocks with high speed and high purity. Deprotection of the glycopeptide intermediate and subsequent glycosyltransferase-catalyzed sugar elongations were performed for generation of the additional diversities with the sugar moieties of glycopeptides using beta1,4-galactosyltransferase (beta1,4-GalT) and two kinds of alpha2,3-sialyltransferases [ST3Gal III; alpha2,3-(N)-SiaT and ST3Gal II; alpha2,3-(O)-SiaT]. These reactions proceeded successfully in the presence of 0.2% Triton X-100 to convert the chemically synthesized trisaccharide glycans to disialylated hexasaccharide.
采用微波辅助固相合成（MA-SPGS）与酶促糖基延伸相结合的新策略，合成了一种具有五个核心-2六糖分支的MUC1相关糖肽（C330H527N46O207，分子量 = 8450.9）。使用糖基化氨基酸构建块，在聚乙二醇-聚（N，N-二甲基丙烯酰胺）共聚物（PEGA）树脂与MA-SPGS的组合中，能以高速和高纯度最佳地实现关键糖肽中间体的合成。对糖肽中间体进行脱保护，随后使用β1,4-半乳糖基转移酶（β1,4-GalT）和两种α2,3-唾液酸转移酶[ST3Gal III；α2,3-（N）-SiaT和ST3Gal II；α2,3-（O）-SiaT]进行糖基转移酶催化的糖基延伸反应，以利用糖肽的糖部分产生更多样性。在0.2% Triton X-100存在的情况下，这些反应成功进行，将化学合成的三糖聚糖转化为双唾液酸化六糖。
