层粘连蛋白-511 和纤维连接蛋白的降解与假丝酵母。

Laminin-511 and fibronectin degradation with Candida yeast.

机构信息

Institute of Dentistry, PB 63, FIN-00014, University of Helsinki, Helsinki, Finland.

出版信息

J Oral Pathol Med. 2009 Nov;38(10):768-72. doi: 10.1111/j.1600-0714.2009.00785.x. Epub 2009 May 21.

DOI:10.1111/j.1600-0714.2009.00785.x

PMID:19486268

Abstract

BACKGROUND

The invasion mechanism of Candida yeast is still partly unknown. In this study, we tested the ability of different commensal Candida yeast to degrade two basement membrane and extracellular matrix proteins: laminin-511 (Lm-511) and plasma fibronectin.

METHODS

Human Lm-511 was produced by an immortal keratinocyte cell line, labelled with (35)S-methionine and immunoprecipitated from the growth medium with monoclonal antibodies. Human plasma fibronectin was purified from plasma samples of blood donors. Sonicated yeast cells and concentrated yeast cell growth media were incubated with Lm-511 in different pH values and the degradation was detected by fluorography. Fibronectin degradation by yeast was visualized by sodium dodecyl-sulphate polyacrylamide gel electrophoresis.

RESULTS

The reduced 220 kDa fibronectin monomers were found to be degraded at pH 7.8 by 10x concentrated growth media of most strains tested and at pH 3.0 the degradation was more pronounced. Sonicated cell fractions of C. tropicalis and C. parapsilosis caused degradation of plasma fibronectin at pH 7.8. Instead, none of the tested Candida cell fractions degraded Lm-511 under these conditions.

CONCLUSIONS

It seems that cleavage of different laminin isoforms by Candida yeast is a laminin-specific process. The ability to cleave human plasma fibronectin is species- and pH-dependent but not hyphal-dependent and also this degradation may affect epithelial integrity.

摘要

背景

白色念珠菌酵母的侵袭机制尚不完全清楚。在这项研究中，我们测试了不同共生白色念珠菌酵母降解两种基底膜和细胞外基质蛋白的能力：层粘连蛋白-511（Lm-511）和血浆纤维连接蛋白。

方法

用人永生化角质形成细胞系产生 Lm-511，用（35）S-甲硫氨酸标记，并用单克隆抗体从生长培养基中免疫沉淀。人血浆纤维连接蛋白从献血者的血浆样本中纯化。用超声处理的酵母细胞和浓缩酵母细胞生长培养基在不同 pH 值下与 Lm-511 孵育，并通过荧光成像检测降解。酵母对纤维连接蛋白的降解通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳可视化。

结果

在 pH 值为 7.8 时，发现大多数测试菌株的 10 倍浓缩生长培养基可降解 220 kDa 的还原型纤维连接蛋白单体，而在 pH 值为 3.0 时，降解更为明显。在 pH 值为 7.8 时，C. tropicalis 和 C. parapsilosis 的超声细胞部分可导致血浆纤维连接蛋白的降解。相比之下，在这些条件下，没有一种测试的白色念珠菌细胞部分可降解 Lm-511。