Characterization and functional analysis of serine proteinase and serine proteinase homologue from the swimming crab Portunus trituberculatus.
Characterization and functional analysis of serine proteinase and serine proteinase homologue from the swimming crab Portunus trituberculatus.
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DOI：
10.1016/j.fsi.2013.04.024
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发表时间：
2013-08
影响因子：
4.7
通讯作者：
Chengwen Song;Z. Cui;Yuan Liu;Qianqian Li;Xihong Li;Guohui Shi;Chunlin Wang
中科院分区：
文献类型：
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作者：
Chengwen Song;Z. Cui;Yuan Liu;Qianqian Li;Xihong Li;Guohui Shi;Chunlin Wang
丝氨酸蛋白酶（SP）及其同源物（SPH）是一类多功能蛋白，在天然免疫系统中起着重要作用。在我们的本研究中，我们进行了适当的校正：丝氨酸蛋白酶同源物PtcSPH（Li等人，[1])从梭子蟹Portunus trituberculatus获得的蛋白酶实际上是丝氨酸蛋白酶，并重新命名为PtcSP。序列分析显示PtcSP和PtSP（Li等人，[2])可能由相同的基因组位点编码，并通过前体mRNA的选择性剪接产生。在PtcSP基因组DNA序列中鉴定出8个外显子。结合我们以前的报道进行了全面的系统发育分析（Cui等人，[3]; Li等人，[1]，[2]）。结果表明，三疣梭子蟹的SP和SPH在基因进化上有不同的起源。为了进一步表征蛋白质的功能，测定重组丝氨酸蛋白酶或同源物的各种生物学功能：蛋白酶活性、抗微生物活性和微生物结合活性。重组蛋白PtcSP具有胰蛋白酶样蛋白酶活性和抑菌活性。PtSPH 1（Li等人，[2])缺乏蛋白水解活性，但显示出对酵母和螃蟹病原菌溶藻弧菌的结合活性。此外，PtcSP的N-末端剪切结构域具有抗菌活性，C-末端SP样结构域具有胰蛋白酶样蛋白酶活性。
Serine proteases (SPs), with their homologues (SPHs), a family of multifunctional proteins, play a crucial role in innate immune system. In our present study, we made an appropriate correction: serine protease homologue PtcSPH (Li et al., [1]) obtained from the swimming crab Portunus trituberculatus was actually a serine protease and re-designated as PtcSP. Sequence analysis revealed PtcSP and PtSP (Li et al., [2]) might be encoded by the same genomic locus and generated by alternative splicing of the pre-mRNA. Eight exons were identified in genomic DNA sequence of PtcSP. A comprehensive phylogenetic analysis was made combined with our previous reports (Cui et al., [3]; Li et al., [1], [2]). The result showed SPs and SPHs ofP. trituberculatushad different origins in gene evolution. To further characterize the function(s) of proteins, the recombinant serine proteases or homologues were assayed for various biological functions: proteinase activity, antimicrobial activity and microorganisms binding activity. The recombinant protein PtcSP exhibited trypsin-like protease activity and antibacterial activity. PtSPH1 (Li et al., [2]) lacked proteolytic activity but displayed binding activity to yeast and the crab pathogenic bacterium, Vibrio alginolyticus. Further, the N-terminal clip domain of PtcSP had antibacterial activity and the C-terminal SP-like domain had trypsin-like protease activity.