The PE-PGRS glycine-rich proteins of Mycobacterium tuberculosis:: a new family of fibronectin-binding proteins?
The PE-PGRS glycine-rich proteins of Mycobacterium tuberculosis:: a new family of fibronectin-binding proteins?
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DOI：
10.1099/00221287-145-12-3487
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发表时间：
1999-12-01
期刊：
影响因子：
--
通讯作者：
Moreno, C
中科院分区：
文献类型：
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作者：
Espitia, C;Laclette, JP;Moreno, C
从结核分枝杆菌粘粒文库中筛选出一个克隆，该克隆是用一种从先前报道的富含脯氨酸蛋白的N-末端序列设计的寡核苷酸筛选的。对4481 bp插入片段的表征表明存在多态性CG重复序列（PGRS），其ORF为2.7 kb，编码81.3 kDa蛋白（PE-PGRS 81）。Southern杂交分析和BLAST-p搜索结果显示，在M.推导的氨基酸序列与GenBank数据库中PE-PGRS家族的几个成员的N-末端约98个残基高度相似，包括与orf 3 '编码的潜在蛋白的部分氨基酸序列以及与Rv 0278 c序列的100%同源性。相邻相关感染单位对数据库中可用的99个PE-PGRS序列的分析表明，临床科学中心PE-PGRS 81被包括在一个组中，其中其最接近的亲属是序列orf 3 '，Rv 0278 c，Rv 0279 c，Rv 1759 c，Rv 3652和Rv 0747。用PE-PGRS 81和Rv 1759 c的完整编码区对结核分枝杆菌、结核分枝杆菌H37 Rv、牛分枝杆菌BCG和分枝杆菌M.结核病临床分离株，表现出复杂的杂交模式的所有菌株。这表明存在菌株内PGRS变异性，与其他PGRS成员报告的一致。相反，用Rv 1759 c的短保守N-末端区域探测将杂交减少到单一条带。该标记物允许鉴定M。缺乏Rv 1759 c的结核病临床菌株。Rv 1759 c的重组C-末端片段显示纤连蛋白结合特性，并被结核分枝杆菌感染患者的血清所识别，表明至少该PE-PGRS成员在结核感染中表达。
A clone was isolated by screening of a cosmid library of Mycobacterium tuberculosis with an oligonucleotide designed from the N-terminal sequence of a previously reported proline-rich protein. Characterization of the 4481 bp insert showed the presence of polymorphic CG-repetitive sequences (PGRSs) with an ORF of 2.7 kb, encoding a 81.3 kDa protein (PE-PGRS81). Southern blot analysis and BLAST-p searches revealed several homologous sequences in the genome of M. tuberculosis, The deduced amino acid sequence was highly similar to a stretch of about 98 residues in the N-terminus present in several members of the PE-PGRS family available in the GenBank database, including 100% identity with the partial amino acid sequence of the potential protein encoded by orf3' as well as with the Rv0278c sequence. A neighbour-joining Related infections Unit, analysis of the 99 PE-PGRS sequences available in the database indicated that Clinical Sciences Centre, PE-PGRS81 is included in a group where its closest relatives are the sequences orf 3', Rv0278c, Rv0279c, Rv1759c, Rv3652 and Rv0747. Probing with the complete coding regions of PE-PGRS81 and Rv1759c in Southern blot assays, on samples of genomic DNA from M, tuberculosis H37Rv, Mycobacterium bovis BCG and M. tuberculosis clinical isolates, showed a complex hybridization pattern for all strains. This shows the existence of intrastrain PGRS variability as reported for other PGRS members. In contrast, probing with the short conserved N-terminal region of Rv1759c reduced the hybridization to a single band. This marker allowed identification of M. tuberculosis clinical strains that lack Rv1759c. A recombinant C-terminal fragment of Rv1759c showed fibronectin-binding properties and was recognized by sera from patients infected with M, tuberculosis, suggesting that at least this member of the PE-PGRS is expressed in tuberculosis infection.