胸膜肺炎放线杆菌金属蛋白酶：克隆与体内表达

Actinobacillus pleuropneumoniae metalloprotease: cloning and in vivo expression.

作者信息

García González Octavio, García Rosa M, de la Garza Mireya, Vaca Sergio, Paniagua Gloria Luz, Mejía Ricardo, Tenorio Víctor R, Negrete-Abascal Erasmo

机构信息

Carrera de Biología, Facultad de Estudios Superiores Iztacala, UNAM, Av. de los Barrios #1, Los Reyes Iztacala, Tlalnepantla, Estado de México 54090, Mexico.

出版信息

FEMS Microbiol Lett. 2004 May 1;234(1):81-6. doi: 10.1016/j.femsle.2004.03.012.

DOI:10.1016/j.femsle.2004.03.012

PMID:15109723

Abstract

The complete amino acid and nucleotide sequence of a secreted metalloprotease produced by Actinobacillus pleuropneumoniae serotype 1 is reported. A clone showing proteolytic activity in cell-free culture media was selected from a genomic library of A. pleuropneumoniae serotype 1 in pUC 19. The sequence obtained contained an open reading frame encoding a protein with 869 amino acids. This protein was identified as a zinc neutral-metalloprotease belonging to the aminopeptidase family, with a predicted molecular weight of approximately 101 kDa. This sequence showed high homology with other predicted or sequenced aminopeptidases reported for different Gram-negative bacteria. Expression of the protease was observed in lung tissue from pigs that died of porcine pleuropneumonia suggesting a role in pathogenesis.

摘要

报道了胸膜肺炎放线杆菌1型分泌的金属蛋白酶的完整氨基酸和核苷酸序列。从胸膜肺炎放线杆菌1型在pUC 19中的基因组文库中筛选出一个在无细胞培养基中显示蛋白水解活性的克隆。获得的序列包含一个编码869个氨基酸的蛋白质的开放阅读框。该蛋白质被鉴定为属于氨肽酶家族的锌中性金属蛋白酶，预测分子量约为101 kDa。该序列与报道的其他不同革兰氏阴性菌的预测或测序氨肽酶具有高度同源性。在死于猪胸膜肺炎的猪的肺组织中观察到该蛋白酶的表达，提示其在发病机制中起作用。