Affinity proteomic approach for identification of an IgA-like protein in Litopenaeus vannamei and study on its agglutination characterization.
An unknown protein reacted with anti-human IgA, namely, IgA-like protein, has been reported in shrimp, but information regarding its identification is not available. In the present study, an affinity proteomic strategy was applied to identify the IgA-like protein of shrimp Litopenaeus vannamei. The protein of 75 kDa was isolated and confirmed by affinity chromatography and Western blotting with goat anti-human IgA, respectively, and then identified as hemocyanin, a member of IgSF, by mass spectrometry. Moreover, our results showed that human IgA and L. vannamei hemocyanin could separately react with goat anti-human IgA or rabbit anti-shrimp affinity hemocyanin (a-hemocyanin). Further evidences indicated that the recombinant protein of the Ig-like conserved domain could react with anti-human IgA. Interestingly, our results indicated that L. vannamei hemocyanin could aggregate with eight species of shrimp pathogenic bacteria and four types of animal erythrocytes directly. These results indicate that L. vannamei hemocyanin, an IgA-like protein, has dual function of reaction with anti-human IgA as an antigen and of activity binding to bacteria and animal erythrocytes as an agglutinin, suggesting its characteristic role as an IgSF molecule. In addition, our approach suggests that affinity proteomics based on heterogeneous antibody can speed up the identification of Fossman antigens.
Zhang
Yueling
Y
Center for Proteomics and Department of Biology, School of Life Sciences, Xiamen University, Xiamen 361005, China.
Wang
Sanying
S
Xu
Anlong
A
Chen
Jun
J
Lin
Bokun
B
Peng
Xuanxian
X
D000818:Animals; D001798:Blood Proteins / Q000032:analysis; D015153:Blotting, Western; D002645:Chickens; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000378:metabolism; D005399:Fishes; D006384:Hemagglutination; D006385:Hemagglutination Inhibition Tests; D006433:Hemocyanins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006458:Hemolymph / Q000737:chemistry; D006801:Humans; D007070:Immunoglobulin A / Q000737:chemistry*; D051379:Mice; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid / Q000494:pharmacology; D033561:Penaeidae / Q000737:chemistry* / Q000382:microbiology; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017422:Sequence Analysis, DNA; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D044149:Vibrio alginolyticus / Q000472:pathogenicity
D001798:Blood Proteins; D007070:Immunoglobulin A; D011506:Proteins; D011994:Recombinant Proteins; D006433:Hemocyanins; D019158:N-Acetylneuraminic Acid