title: Antibacterial activity of serine protease inhibitor 1 from kuruma shrimp Marsupenaeus japonicus.
abstract: Serine protease inhibitors (Serpins) are a large family of protease inhibitors involved in many critical biological processes such as blood coagulation, fibrinolysis, programmed cell death, development, and innate immunity. We identified MjSerp1, a serpin in the kuruma shrimp Marsupenaeus japonicus. The MjSerp1 cDNA has a 1239 bp open reading frame (ORF) that encodes a 412-amino acid protein with a 23 aa signal peptide and a classic serpin domain. MjSerp1 has a calculated molecular mass of 46.3 kDa and a predicted isoelectric point of 5.51. MjSerp1 is mainly expressed in the hepatopancreas and the intestines, and is moderately expressed in hemocytes. Expression pattern analysis indicated that MjSerp1 is upregulated in the hepatopancreas after Vibrio anguillarum challenge. rMjSerp1 inhibits three Gram-positive bacteria and two Gram-negative bacteria, but does not inhibit phenoloxidase activity. The microorganism binding assay showed that rMjSerp1 closely binds to both Gram-positive and Gram-negative bacteria. MjSerp1 also exhibits inhibitory activity against microbial serine proteases, such as subtilisin A and proteinase K, indicating that MjSerp1 acts as a microbial serine protease inhibitor. rMjSerp1 injection into shrimp enhances V. anguillarum clearance, but MjSerp1 knockdown through RNA interference impairs Vibrio clearance in vivo. These results indicate that MjSerp1 functions as a direct effector in the bacterial clearance of M. japonicus. All together, our findings provide novel evidences for the serine protease inhibitor in shrimp immunity.
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Yan-Ran
authors.initials: YR
authors.lastname: Zhao
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Yi-Hui
authors.initials: YH
authors.lastname: Xu
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Hai-Shan
authors.initials: HS
authors.lastname: Jiang
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Xu
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Xiao-Fan
authors.initials: XF
authors.lastname: Zhao
authors.affiliation: MOE Key Laboratory of Plant Cell Engineering and Germplasm Innovation, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China. Electronic address: jxwang@sdu.edu.cn.
authors.forename: Jin-Xing
authors.initials: JX
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000378:metabolism*; D001156:Artemia / Q000276:immunology*; D060829:Arthropod Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D019286:Endopeptidase K / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000276:immunology*; D016908:Gram-Positive Bacterial Infections / Q000276:immunology*; D043143:Hepatopancreas / Q000378:metabolism*; D007113:Immunity, Innate; D007413:Intestinal Mucosa / Q000378:metabolism*; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015843:Serpins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013381:Subtilisins / Q000378:metabolism; D059467:Transcriptome; D015854:Up-Regulation; D014733:Vibrio / Q000276:immunology*; D014735:Vibrio Infections / Q000276:immunology*
chemical_list: D000900:Anti-Bacterial Agents; D060829:Arthropod Proteins; D001426:Bacterial Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D013381:Subtilisins; D019286:Endopeptidase K
keywords: Antibacterial activity; Innate immunity; Marsupenaeus japonicus; Serpins