title: A double WAP domain-containing protein Es-DWD1 from Eriocheir sinensis exhibits antimicrobial and proteinase inhibitory activities.
abstract: Whey acidic proteins (WAP) belong to a large gene family of antibacterial peptides, which are critical in the host immune response against microbial invasion. The common feature of these proteins is a single WAP domain maintained by at least one four-disulfide core (4-DSC) structure rich in cysteine residues. In this study, a double WAP domain (DWD)-containing protein, Es-DWD1, was first cloned from the Chinese mitten crab (Eriocheirsinensis). The full-length Es-DWD1cDNA was 1193 bp, including a 411 bp open reading frame (ORF) encoding 136 amino acids with a signal peptide of 22 amino acids in the N-terminus. A comparison with other reported invertebrate and vertebrate sequences revealed the presence of WAP domains characteristic of WAP superfamilies. As determined by quantitative real-time RT-PCR, Es-DWD1 transcripts were ubiquitously expressed in all tissues, but it was up-regulated in hemocytes post-challenge with pathogen-associated molecular patterns (PAMPs). The mature recombinant Es-DWD1 (rEs-DWD1) protein exhibited different binding activities to bacteria and fungus. Moreover, rEs-DWD1 could exert agglutination activities against Bacillus subtilis and Pichiapastoris and demonstrated inhibitory activities against the growth of Staphylococcus aureus, Aeromonas hydrophila and P. pastoris. Furthermore, rEs-DWD1 showed a specific protease inhibitory activity in B. subtilis. Coating of rEs-DWD1 onto agarose beads enhanced encapsulation of the beads by crab hemocytes. Collectively, the results suggest that Es-DWD1 is a double WAP domain containing protein with antimicrobial and proteinase inhibitory activities, which play significant roles in the immunity of crustaceans.
authors: School of Life Science, East China Normal University, Shanghai, China.
authors: Shuang
authors: S
authors: Li
authors: Xing-Kun
authors: XK
authors: Jin
authors: Xiao-Nv
authors: XN
authors: Guo
authors: Ai-Qing
authors: AQ
authors: Yu
authors: Min-Hao
authors: MH
authors: Wu
authors: Shang-Jian
authors: SJ
authors: Tan
authors: You-Ting
authors: YT
authors: Zhu
authors: Wei-Wei
authors: WW
authors: Li
authors: Qun
authors: Q
authors: Wang
mesh_terms: D000372:Agglutination Tests; D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D001483:Base Sequence; D003386:Brachyura / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D010802:Phylogeny; D011480:Protease Inhibitors / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D011480:Protease Inhibitors; D011994:Recombinant Proteins; C570318:double WAP domain-containing protein, Eriocheir sinensis
references: Annu Rev Immunol. 1995;13:61-92
references: 7612236
references: J Biol Chem. 1990 Apr 25;265(12):6854-9
references: 2324101
references: Biochem Soc Trans. 2011 Oct;39(5):1403-8
references: 21936823
references: J Mol Graph Model. 1999 Apr;17(2):106-13, 134-6
references: 10680116
references: Cancer Res. 1988 Sep 15;48(18):5203-9
references: 3136918
references: Biol Reprod. 2004 Nov;71(5):1484-90
references: 15229136
references: Fish Shellfish Immunol. 2011 Jul;31(1):142-7
references: 21549196
references: Dev Comp Immunol. 2008;32(4):317-29
references: 17706773
references: Methods. 2001 Dec;25(4):402-8
references: 11846609
references: Biochem Biophys Res Commun. 1989 Oct 16;164(1):496-502
references: 2553020
references: PLoS One. 2010 Jun 17;5(6):e11175
references: 20567508
references: Dev Comp Immunol. 2010 Jul;34(7):734-40
references: 20144896
references: Comp Biochem Physiol B Biochem Mol Biol. 1995 Oct;112(2):385-91
references: 7584865
references: Immunol Rev. 2004 Apr;198:169-84
references: 15199962
references: J Biol Chem. 1990 Sep 5;265(25):14791-5
references: 2394696
references: Eur J Biochem. 1994 Jul 15;223(2):389-94
references: 8055907
references: Comp Biochem Physiol B Biochem Mol Biol. 2009 Oct;154(2):203-10
references: 19545640
references: Mol Immunol. 2006 Mar;43(9):1490-6
references: 16144710
references: J Cell Sci. 1996 Aug;109 ( Pt 8):2053-60
references: 8856501
references: Fish Shellfish Immunol. 2010 Jan;28(1):134-42
references: 19840856
references: Res Microbiol. 2004 Oct;155(8):630-5
references: 15380550
references: Dev Comp Immunol. 2006;30(11):998-1008
references: 16519941
references: Fish Shellfish Immunol. 2009 Dec;27(6):739-47
references: 19766195
references: Dev Comp Immunol. 1983 Spring;7(2):229-39
references: 6409683
references: PLoS One. 2012;7(3):e33735
references: 22442720
references: Dev Comp Immunol. 2008;32(12):1497-509
references: 18602420
references: Infect Immun. 1996 Nov;64(11):4520-4
references: 8890201
references: J Invertebr Pathol. 2008 Oct;99(2):141-5
references: 18501924
references: Dev Comp Immunol. 2010 Dec;34(12):1274-82
references: 20638410
references: Mol Immunol. 2007 Feb;44(6):1065-74
references: 16982096
references: Mol Immunol. 2006 Feb;43(4):388-94
references: 16310052
references: Dev Comp Immunol. 2009 Apr;33(4):583-91
references: 19041340
references: J Bacteriol. 1953 Jul;66(1):1-5
references: 13069457
references: Comp Biochem Physiol C Toxicol Pharmacol. 2003 Apr;134(4):431-8
references: 12727292
references: Fish Shellfish Immunol. 2004 May;16(5):599-611
references: 15110334
references: J Biotechnol. 2007 Jan 20;127(4):605-14
references: 16987562
references: Dev Comp Immunol. 2012 Jan;36(1):208-15
references: 21855569
references: Fish Shellfish Immunol. 2008 Dec;25(6):775-81
references: 18974012
references: J Invertebr Pathol. 2004 May-Jun;86(1-2):59-60
references: 15145254
references: Biochem Soc Trans. 2002 Apr;30(2):111-5
references: 12023836
references: J Infect Dis. 1997 Sep;176(3):740-7
references: 9291323
references: FEBS Lett. 1989 Aug 28;254(1-2):111-4
references: 2476331
references: J Immunol. 2007 Dec 15;179(12):8425-34
references: 18056389
references: Mar Biotechnol (NY). 2002 Oct;4(5):487-94
references: 14961242
references: Comp Biochem Physiol B Biochem Mol Biol. 2004 Aug;138(4):415-22
references: 15325343
references: Mol Immunol. 2008 Feb;45(4):1085-93
references: 17850873
references: Dev Comp Immunol. 2012 Jan;36(1):183-90
references: 21798281
references: Adv Exp Med Biol. 2001;484:95-112
references: 11419011
references: Dev Comp Immunol. 2005;29(5):385-91
references: 15707660
references: Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6682-7
references: 9192625
references: Dev Comp Immunol. 2008;32(7):758-72
references: 18222540
references: Gene. 2001 Jan 24;263(1-2):121-30
references: 11223250
references: J Immunol. 2003 Feb 15;170(4):1973-9
references: 12574366
references: Fish Shellfish Immunol. 2007 Nov;23(5):1119-26
references: 17664073
references: J Bacteriol. 1946 Feb;51(2):149-67
references: 16561064
references: Nucleic Acids Res. 1982 Apr 24;10(8):2677-84
references: 6896234