Aebi C, Maciver I, Latimer J L, Cope L D, Stevens M K, Thomas S E, McCracken G H, Hansen E J
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235-9048, USA.
Infect Immun. 1997 Nov;65(11):4367-77. doi: 10.1128/iai.65.11.4367-4377.1997.
The high-molecular-weight UspA protein of Moraxella catarrhalis has been described as being both present on the surface of all M. catarrhalis disease isolates examined to date and a target for a monoclonal antibody (MAb 17C7) which enhanced pulmonary clearance of this organism in a mouse model system (M. E. Helminen et al., J. Infect. Dis. 170:867-872, 1994). A recombinant bacteriophage that formed plaques which bound MAb 17C7 was shown to contain a M. catarrhalis gene, designated uspA1, that encoded a protein with a calculated molecular weight of 88,271. Characterization of an isogenic uspA1 mutant revealed that elimination of expression of UspA1 did not eliminate the reactivity of M. catarrhalis with MAb 17C7. In addition, N-terminal amino acid analysis of internal peptides derived from native UspA protein and Southern blot analysis of M. catarrhalis chromosomal DNA suggested the existence of a second UspA-like protein. A combination of epitope mapping and ligation-based PCR methods identified a second M. catarrhalis gene, designated uspA2, which also encoded the MAb 17C7-reactive epitope. The UspA2 protein had a calculated molecular weight of 62,483. Both the isogenic uspA1 mutant and an isogenic uspA2 mutant possessed the ability to express a very-high-molecular-weight antigen that bound MAb 17C7. Southern blot analysis indicated that disease isolates of M. catarrhalis likely possess both uspA1 and uspA2 genes. Both UspA1 and UspA2 most closely resembled adhesins produced by other bacterial pathogens.
卡他莫拉菌的高分子量UspA蛋白已被描述为存在于迄今为止检测的所有卡他莫拉菌疾病分离株的表面,并且是一种单克隆抗体(MAb 17C7)的靶标,该抗体在小鼠模型系统中增强了这种生物体的肺部清除能力(M. E. Helminen等人,《传染病杂志》170:867 - 872,1994)。一种形成能结合MAb 17C7噬菌斑的重组噬菌体被证明含有一个卡他莫拉菌基因,命名为uspA1,该基因编码一种计算分子量为88,271的蛋白质。对一个同基因uspA1突变体的表征显示,UspA1表达的消除并未消除卡他莫拉菌与MAb 17C7的反应性。此外,对源自天然UspA蛋白的内部肽段进行的N端氨基酸分析以及对卡他莫拉菌染色体DNA的Southern印迹分析表明存在第二种UspA样蛋白。表位作图和基于连接的PCR方法相结合,鉴定出第二个卡他莫拉菌基因,命名为uspA2,它也编码MAb 17C7反应性表位。UspA2蛋白的计算分子量为62,483。同基因uspA1突变体和同基因uspA2突变体都具有表达一种能结合MAb 17C7的非常高分子量抗原的能力。Southern印迹分析表明,卡他莫拉菌的疾病分离株可能同时拥有uspA1和uspA2基因。UspA1和UspA2与其他细菌病原体产生的黏附素最为相似。