Identification and functional characterizations of serpin8, a potential prophenoloxidase-activating protease inhibitor in Pacific white shrimp, Litopenaeus vannamei.
Serpins have been characterized from varieties of organisms by their inhibitory roles on serine or cysteine proteases. However, research for the functional study of serpins in crustacean is relatively small. To fully clarify the immune characterizations of serpin, a novel serpin (named Lvserpin8) encoding 414 amino acids with a 19-amino acid signal peptide and a serpin domain was identified from the Pacific white shrimp Litopenaeus vannamei. Sequence analysis indicated that the genomic Lvserpin8 gene contains 5 exons and 4 introns. The P1 residues of the predicted scissile bond in the reactive center loop (RCL) region represented for Lysine (Lys), which is in accordance with Pmserpin8, Dmserpin27A, Ofserpin3, Bmserpin3 and Msserpin3. Quantitative results showed that high mRNA expression of Lvserpin8 was detected in hepatopancreas and testis. Notably, a significant increase of Lvserpin8 was appeared post injection of Vibrio anguillarum, and Micrococcus lysodeikticus. Moreover, Lvserpin8 was knocked down in vivo by double-stranded RNA (dsRNA) mediated RNA interference (RNAi). Suppression of Lvserpin8 led to a significant increase in the transcripts of LvPPAE2 (Prophenoloxidase-activating Enzyme 2) and cumulative mortality. What's more, recombinant Lvserpin8 protein (rLvserpin8) displayed inhibition roles on trypsin activity, and prophenoloxidase activation. Taken together, the results implied that Lvserpin8 may be involved in shrimp innate immunity via the inhibition of prophenoloxidase-activating proteases.
College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling 712100, China.
Yongjie
Y
Liu
College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling 712100, China.
Yuhang
Y
Sun
College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling 712100, China.
Qiai
Q
Wang
College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling 712100, China.
Fujun
F
Hou
College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling 712100, China. Electronic address: liuxiaolin@nwsuaf.edu.cn.
Xiaolin
X
Liu
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D007113:Immunity, Innate; D008837:Micrococcus / Q000502:physiology; D009928:Organ Specificity; D033561:Penaeidae / Q000145:classification / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology; D010802:Phylogeny; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015843:Serpins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D014733:Vibrio / Q000502:physiology
D060829:Arthropod Proteins; D018076:DNA, Complementary; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015843:Serpins
Litopenaeus vannamei; Prophenoloxidase system; Proteases; Serpin8; dsRNA