Hamil Katherine G, Liu Qiang, Sivashanmugam P, Yenugu Suresh, Soundararajan Rama, Grossman Gail, Richardson Richard T, Zhang Yong-Lian, O'Rand Michael G, Petrusz Peter, French Frank S, Hall Susan H
Department of Pediatrics, University of North Carolina School of Medicine, Chapel Hill 27599, USA.
Endocrinology. 2002 Jul;143(7):2787-96. doi: 10.1210/endo.143.7.8925.
Cystatin (CST)11, a novel member of the CST type 2 family of cysteine protease inhibitors, was identified in Macaca mulatta epididymis by subtractive hybridization cloning. The human CST11 gene on chromosome 20p11.2 is located near three other CST genes expressed predominantly in the male reproductive tract. The CST11 gene spans three exons, a structure similar to that of other CST family 2 genes. An exon 2-deleted alternative transcript (CST11Delta2) was also identified. CST11 mRNA is expressed only in the epididymis as judged by Northern blot hybridization and is androgen regulated. The protein is most abundant in the initial segment, but is detected throughout the epididymis and on ejaculated human sperm. The calculated tertiary structure of CST11 reveals that the three regions corresponding to the protease inhibitory wedge of CST3 are similarly juxtaposed in CST11, consistent with protease inhibitor function. Intact and exon 2-deleted CST11 recombinant proteins were tested for antibacterial activity. After a 2-h incubation of Escherichia coli with 50 microg/ml recombinant CST11 or CST11Delta2, bacterial colony-forming units were reduced to 30% of control, indicating that both forms have antimicrobial activity.
胱抑素(CST)11是半胱氨酸蛋白酶抑制剂CST 2家族的一个新成员,通过消减杂交克隆在猕猴附睾中被鉴定出来。位于20p11.2染色体上的人类CST11基因,靠近其他三个主要在男性生殖道表达的CST基因。CST11基因跨越三个外显子,其结构与其他CST家族2基因相似。还鉴定出一种缺失外显子2的可变转录本(CST11Delta2)。通过Northern印迹杂交判断,CST11 mRNA仅在附睾中表达,且受雄激素调节。该蛋白在起始段最为丰富,但在整个附睾以及射出的人类精子上均能检测到。CST11的计算三级结构显示,与CST3的蛋白酶抑制楔形相对应的三个区域在CST11中也类似地并列,这与蛋白酶抑制剂功能一致。对完整的和缺失外显子2的CST11重组蛋白进行了抗菌活性测试。用50微克/毫升的重组CST11或CST11Delta2孵育大肠杆菌2小时后,细菌集落形成单位降至对照的30%,表明这两种形式都具有抗菌活性。