Stockbauer K E, Magoun L, Liu M, Burns E H, Gubba S, Renish S, Pan X, Bodary S C, Baker E, Coburn J, Leong J M, Musser J M
Institute for the Study of Human Bacterial Pathogenesis, Department of Pathology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Proc Natl Acad Sci U S A. 1999 Jan 5;96(1):242-7. doi: 10.1073/pnas.96.1.242.
The human pathogenic bacterium group A Streptococcus produces an extracellular cysteine protease [streptococcal pyrogenic exotoxin B (SpeB)] that is a critical virulence factor for invasive disease episodes. Sequence analysis of the speB gene from 200 group A Streptococcus isolates collected worldwide identified three main mature SpeB (mSpeB) variants. One of these variants (mSpeB2) contains an Arg-Gly-Asp (RGD) sequence, a tripeptide motif that is commonly recognized by integrin receptors. mSpeB2 is made by all isolates of the unusually virulent serotype M1 and several other geographically widespread clones that frequently cause invasive infections. Only the mSpeB2 variant bound to transfected cells expressing integrin alphavbeta3 (also known as the vitronectin receptor) or alphaIIbbeta3 (platelet glycoprotein IIb-IIIa), and binding was blocked by a mAb that recognizes the streptococcal protease RGD motif region. In addition, mSpeB2 bound purified platelet integrin alphaIIbbeta3. Defined beta3 mutants that are altered for fibrinogen binding were defective for SpeB binding. Synthetic peptides with the mSpeB2 RGD motif, but not the RSD sequence present in other mSpeB variants, blocked binding of mSpeB2 to transfected cells expressing alphavbeta3 and caused detachment of cultured human umbilical vein endothelial cells. The results (i) identify a Gram-positive virulence factor that directly binds integrins, (ii) identify naturally occurring variants of a documented Gram-positive virulence factor with biomedically relevant differences in their interactions with host cells, and (iii) add to the theme that subtle natural variation in microbial virulence factor structure alters the character of host-pathogen interactions.
人类致病细菌A群链球菌产生一种细胞外半胱氨酸蛋白酶[链球菌致热外毒素B(SpeB)],它是侵袭性疾病发作的关键毒力因子。对全球收集的200株A群链球菌分离株的speB基因进行序列分析,确定了三种主要的成熟SpeB(mSpeB)变体。其中一种变体(mSpeB2)包含一个精氨酸 - 甘氨酸 - 天冬氨酸(RGD)序列,这是一种通常被整合素受体识别的三肽基序。mSpeB2由异常毒力的M1血清型的所有分离株以及其他几个经常引起侵袭性感染的地理分布广泛的克隆产生。只有mSpeB2变体与表达整合素αvβ3(也称为玻连蛋白受体)或αIIbβ3(血小板糖蛋白IIb - IIIa)的转染细胞结合,并且结合被识别链球菌蛋白酶RGD基序区域的单克隆抗体阻断。此外,mSpeB2与纯化的血小板整合素αIIbβ3结合。对纤维蛋白原结合发生改变的特定β3突变体对SpeB结合有缺陷。具有mSpeB2 RGD基序但不具有其他mSpeB变体中存在的RSD序列的合成肽阻断了mSpeB2与表达αvβ3的转染细胞的结合,并导致培养的人脐静脉内皮细胞脱离。这些结果(i)确定了一种直接结合整合素的革兰氏阳性毒力因子,(ii)确定了一种已记录的革兰氏阳性毒力因子的天然变体,它们在与宿主细胞相互作用方面具有生物医学相关差异,并且(iii)进一步证明了微生物毒力因子结构的细微自然变异会改变宿主 - 病原体相互作用的特性这一主题。