Characterization of mycobacterial protein glycosyltransferase activity using synthetic peptide acceptors in a cell-free assay.
Characterization of mycobacterial protein glycosyltransferase activity using synthetic peptide acceptors in a cell-free assay.
复制标题
在无细胞测定中使用合成肽受体表征分枝杆菌蛋白糖基转移酶活性。
DOI：
10.1093/glycob/cwf051
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发表时间：
2002
期刊：
影响因子：
4.3
通讯作者：
Young,Douglas
中科院分区：
文献类型：
--
作者：
Cooper,HowardN;Gurcha,SudagarS;Nigou,Jérôme;Brennan,PatrickJ;Belisle,JohnT;Besra,GurdyalS;Young,Douglas
Synthetic peptides derived from a 45-kDa glycoprotein antigen ofMycobacterium tuberculosiswere shown to function as glycosyltransferase acceptors for mannose residues in a mannosyltransferase cell-free assay. The mannosyltransferase activity was localized within both isolated membranes and a P60 cell wall fraction prepared from the rapidly growing mycobacterial strain,Mycobacterium smegmatis. Incorporation of radiolabel from GDP-[14C]mannose was inhibited by the addition of amphomycin, indicating that the glycosyl donor for the peptide acceptors was a member of the mycobacterial polyprenol-P-mannose (PPM) family of activated glycosyl donors. Furthermore, a direct demonstration of transfer from thein situgenerated PP[14C]Ms was also demonstrated. It was also found that the enzyme activity was sensitive to changes in overall peptide length and amino acid composition. Because glycoproteins are present on the mycobacterial cell surface and are available for interaction with host cells during infection, protein glycosyltransferases may provide novel drug targets. The development of a cell-free mannosyltransferase assay will now facilitate the cloning and biochemical characterisation of the relevant enzymes fromM. tuberculosis.
