Identification of SpyA, a novel ADP-ribosyltransferase of Streptococcus pyogenes
Identification of SpyA, a novel ADP-ribosyltransferase of Streptococcus pyogenes
复制标题
DOI：
10.1111/j.1365-2958.2004.04262.x
复制
发表时间：
2004-10-01
影响因子：
3.6
通讯作者：
Collins, CM
中科院分区：
文献类型：
--
作者：
Coye, LH;Collins, CM
化脓性链球菌是呼吸道和皮肤感染的病原体，会产生大量外毒素来引起感染。该报告鉴定了该生物体产生的一种新外毒素，称为 SpyA，为 S. py 基因 DP-核糖基化毒素。 SpyA，分子量 24.9，与 ADP-核糖转移酶 (ADPRT)、金黄色葡萄球菌 EDIN 和肉毒梭菌 C3 具有氨基酸同一性。重组SpyA能够水解β-NAD(+)，并且该活性依赖于187位处的谷氨酸。SpyA具有推定的双谷氨酸活性位点，并且与大多数双谷氨酸ADPRT类似，能够ADP-核糖基化聚-L-精氨酸。 SpyA 修饰了 CHO 和 HeLa 细胞裂解物中的大量蛋白质。修饰蛋白的二维凝胶分析和 MALDI-TOF MS 分析表明，波形蛋白、原肌球蛋白和肌动蛋白（所有细胞骨架蛋白）都是靶标。 SpyA 在 HeLa 细胞中的表达导致肌动蛋白微丝的损失。我们假设 SpyA 由化脓性链球菌产生，可破坏细胞骨架结构并促进宿主定植。
Streptococcus pyogenes, the aetiological agent of both respiratory and skin infections, produces numerous exotoxins to establish infection. This report identifies a new exotoxin produced by this organism, termed SpyA, for S. py ogenes DP-ribosylating toxin. SpyA, MW 24.9, has amino acid identity with the ADP-riboslytransferases (ADPRTs) Staphylococcus aureus EDIN and Clostridium botulinum C3. Recombinant SpyA was able to hydrolyse beta-NAD(+), and this activity was dependent on a glutamate at position 187. SpyA has a putative biglutamate active site, and similar to most biglutamate ADPRTs, was able to ADP-ribosylate poly-L-arginine. SpyA modified numerous proteins in both CHO and HeLa cell lysates. Two-dimesional gel analysis and MALDI-TOF MS analysis of modified proteins indicated that vimentin, tropomyosin and actin, all cytoskeletal proteins, are targets. Expression of spyA in HeLa cells resulted in loss of actin microfilaments. We hypothesize that SpyA is produced by S. pyogenes to disrupt cytoskeletal structures and promote colonization of the host.