白色念珠菌细胞中β-1,2-甘露糖基转移酶的检测

Detection of beta-1,2-mannosyltransferase in Candida albicans cells.

作者信息

Suzuki A, Takata Y, Oshie A, Tezuka A, Shibata N, Kobayashi H, Okawa Y, Suzuki S

机构信息

Second Department of Hygienic Chemistry, Tohoku College of Pharmacy, Miyagi, Japan.

出版信息

FEBS Lett. 1995 Oct 16;373(3):275-9. doi: 10.1016/0014-5793(95)01061-i.

DOI:10.1016/0014-5793(95)01061-i

PMID:7589482

Abstract

A particulate insoluble fraction from Candida albicans J-1012 (serotype A) strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Man beta 1-->2Man alpha 1-->(2Man alpha 1-->)(2)2Man (alpha beta Man5), in the presence of GDP-mannose followed by high performance liquid chromatography showed the formation of a mannohexaose. Analysis of the product by 1H NMR indicates that alpha beta Man5 was changed to Man beta 1-->2Man beta 1-->2Man alpha 1-->(2Man alpha 1-->)2 2Man (alpha beta Man6). This beta-1,2-mannosyltransferase (ManTase) II activity was completely inhibited by Zn2+ and was not restored by the addition of EDTA. The corresponding enzyme fraction from C. albicans NIH B-792 (serotype B) strain cells, the mannan of which does not possess both the alpha beta Man5 and alpha beta Man6 side chains, also exhibited the same beta-1,2-ManTase II activity.

摘要

白色念珠菌J-1012（A型血清型）菌株细胞的颗粒不溶部分是在用1% Triton X-100提取细胞匀浆105,000 x g沉淀后的残渣。该部分与甘露五糖Manβ1→2Manα1→(2Manα1→)(2)2Man（αβMan5）在GDP-甘露糖存在下孵育，随后进行高效液相色谱分析，结果显示形成了甘露六糖。通过1H NMR对产物进行分析表明，αβMan5转变为Manβ1→2Manβ1→2Manα1→(2Manα1→)2 2Man（αβMan6）。这种β-1,2-甘露糖基转移酶（ManTase）II活性被Zn2+完全抑制，且添加EDTA后无法恢复。来自白色念珠菌NIH B-792（B型血清型）菌株细胞的相应酶部分，其甘露聚糖不具有αβMan5和αβMan6侧链，也表现出相同的β-1,2-ManTase II活性。