Characterization and expression of HLysG2, a basic goose-type lysozyme from the human eye and testis
Characterization and expression of HLysG2, a basic goose-type lysozyme from the human eye and testis
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HLysG2（一种来自人眼和睾丸的碱性鹅型溶菌酶）的表征和表达
DOI：
10.1016/j.molimm.2010.10.008
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发表时间：
2011-01-01
影响因子：
3.6
通讯作者：
Yu, Long
中科院分区：
文献类型：
--
作者：
Huang, Peng;Li, Wen-shu;Yu, Long
溶菌酶通过裂解细菌细胞在人类先天免疫中发挥重要作用。我们描述的第一次，人类溶菌酶G样2（HLysG 2），哺乳动物G型溶菌酶的实际表现。RT-PCR结果显示，HLysG 2基因在眼和睾丸组织中有转录。使用20凝胶电泳从人泪液中检测到一个斑点，并使用MALDI-TOF/TOF MS和MASCOT搜索鉴定为HLysG 2，匹配评分为140，整个氨基酸序列的序列覆盖率为27%。为了深入了解HLysG 2的体外抗菌活性，将成熟肽编码区克隆到毕赤酵母中进行异源表达。重组HLysG 2在pH 6.0和30 ℃下具有最佳活性，在钠离子浓度为75 mM时达到1.2 × 10（4）U/mg的峰值活性，并显示出比人c型溶菌酶（HLysC）更高的耐盐性。重组HlysG 2抑制革兰氏阳性菌生长，不抑制革兰氏阴性菌和白色念珠菌生长。结果表明，HLysG 2是一种有效的抗菌蛋白，可能在人眼的先天免疫中发挥作用。(C)2010爱思唯尔有限公司版权所有。
Lysozyme plays an important role in human innate immunity by causing bacterial cell lysis. We describe for the first time, the actual performance of human lysozyme g-like 2 (HLysG2), a mammalian g-type lysozyme. RT-PCR revealed that the HLysG2 gene was transcribed in eye and testis tissues. A spot was detected from human tears using 20 gel electrophoresis and was identified as HLysG2 using MALDI-TOF/TOF MS and a MASCOT search with a matching score of 140 and 27% sequence coverage of the whole amino acid sequence. To gain insight into the in vitro antimicrobial activities of HLysG2, the mature peptide-coding region was cloned into Pichia pastoris for heterogeneous expression. Recombinant HLysG2, had an optimal at pH 6.0 and 30 degrees C, reached the peak activity of 1.2 x 10(4) U/mg at the sodium ion concentration of 75 mM and showed a higher salt tolerance than human c-type lysozyme (HLysC). Recombinant HlysG2 inhibited Gram-positive bacterial growth and did not inhibit Gram-negative bacterial and Candida albicans growth. Results indicated that HLysG2 is a potent antibacterial protein that may play a role in the innate immunity of the human eye. (C) 2010 Elsevier Ltd. All rights reserved.