MOSPD2 is a receptor mediating the LEAP-2 effect on monocytes/macrophages in a teleost,Boleophthalmus pectinirostris  

作　　者：Chang-Hong Li Jie Chen Li Nie Jiong Chen 

机构地区：[1]State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products,Ningbo University,Ningbo,Zhejiang 315211,China [2]Laboratory of Biochemistry and Molecular Biology,School of Marine Sciences,Meishan Campus,Ningbo University,Ningbo,Zhejiang 315832,China [3]Key Laboratory of Applied Marine Biotechnology of Ministry of Education,Meishan Campus,Ningbo University,Ningbo,Zhejiang 315832,China [4]College of Ecology,Lishui University,Lishui,Zhejiang 323000,China

出　　处：《Zoological Research》2020年第6期644-655,共12页动物学研究（英文）

基　　金：supported by the National Natural Science Foundation of China(31972821,31772876);Zhejiang Provincial Natural Science Foundation of China(LZ18C190001);Scientific Innovation Team Project of Ningbo(2015C110018);K.C.Wong Magna Fund in Ningbo University。

摘　　要：Liver-expressed antimicrobial peptide 2(LEAP-2)is a cationic peptide that plays an important role in a host’s innate immune system.We previously demonstrated that mudskipper(Boleophthalmus pectinirostris)LEAP-2(BpLEAP-2)induces chemotaxis and activation of monocytes/macrophages(MO/MΦ).However,the molecular mechanism by which BpLEAP-2 regulates MO/MΦ remains unclear.In this study,we used yeast twohybrid cDNA library screening to identify mudskipper protein(s)that interacted with BpLEAP-2,and characterized a sequence encoding motile sperm domain-containing protein 2(BpMOSPD2).The interaction between BpLEAP-2 and BpMOSPD2 was subsequently confirmed by co-immunoprecipitation(Co-IP).Sequence analyses revealed that the predicted BpMOSPD2 contained an N-terminal extracellular portion composed of a CRAL-TRIO domain and a motile sperm domain,a C-terminal transmembrane domain,and a short cytoplasmic tail.Phylogenetic tree analysis indicated that BpMOSPD2 grouped tightly with fish MOSPD2 homologs and was most closely related to that of the Nile tilapia(Oreochromis niloticus).The recombinant BpMOSPD2 was produced by prokaryotic expression and the corresponding antibody was prepared for protein concentration determination.RNA interference was used to knockdown BpMOSPD2 expression in the mudskipper MO/MΦ,and the knockdown efficiency was confirmed by quantitative real-time polymerase chain reaction(qRT-PCR)and western blotting.Knockdown of BpMOSPD2 significantly inhibited BpLEAP-2-induced chemotaxis of mudskipper MO/MΦand BpLEAP-2-induced bacterial killing activity.Furthermore,knockdown of BpMOSPD2 inhibited the effect of BpLEAP-2 on mRNA expression levels of BpIL-10,BpTNFα,BpIL-1β,and BpTGFβ in MO/MΦ.In general,BpMOSPD2 directly interacted with BpLEAP-2,and mediated the effects of BpLEAP-2 on chemotaxis and activation of mudskipper MO/MΦ.This is the first identification of MOSPD2 as a receptor for LEAP-2.

关 键 词：LEAP-2 MONOCYTE/MACROPHAGE MOSPD2 RNA interference Yeast two-hybrid 

分 类 号：Q952[生物学—动物学]