title: Yellow head virus binding to cell surface proteins from Penaeus monodon hemocytes.
abstract: Our previous data revealed that viral particles of yellow head virus (YHV) specifically interacted with granule-containing hemocytes. After isolation of targeted hemocytes, biotinylation was performed using Biotin-NSH-LC. Biotinylated protein was extracted and separated by 2-D PAGE. Electro-transferred proteins on a nitrocellulose membrane were probed with streptavidin-HRP complex to detect biotinylated proteins. The data from 2-D PAGE combined with affinity pull down purification revealed 8 and 6 biotinylated proteins specific to hyaline and granule containing hemocytes, respectively. Four proteins were found in common for both two hemocytes. The majority of proteins detected in granular hemocytes are membrane-associated proteins and immune-related proteins such as alpha-2-macroglobulin (A2M), kazal-type serine protease inhibitor (SPI) and crustin. CrustinPm1 was found to bind to YHV as shown with biotinylation pull-down assay and confirmed with two-dimensional virus overlay protein binding assay (2-D VOPBA). The expression of crustinPm1 was observed in semigranular and granular hemocytes whereas very low or no expression occurred in hyaline hemocytes. CrustinPm1 appears to either be directly involved in cellular binding or mediating virus internalization into permissive hemocytes.
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhonpathom, Thailand.
authors.forename: Phattara-Orn
authors.initials: PO
authors.lastname: Havanapan
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhonpathom, Thailand.
authors.forename: Suparat
authors.initials: S
authors.lastname: Taengchaiyaphum
authors.affiliation: Department of Technology and Industries, Faculty of Science and Technology, Prince of Songkla University, Pattani, Thailand.
authors.forename: Apichai
authors.initials: A
authors.lastname: Bourchookarn
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhonpathom, Thailand.
authors.forename: Albert J
authors.initials: AJ
authors.lastname: Ketterman
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhonpathom, Thailand. Electronic address: chartchai.kri@mahidol.ac.th.
authors.forename: Chartchai
authors.initials: C
authors.lastname: Krittanai
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism; D019921:Biotinylation; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005455:Fluorescent Antibody Technique; D006434:Hemocytes / Q000378:metabolism* / Q000821:virology*; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000378:metabolism*; D033561:Penaeidae / Q000821:virology*; D046828:Roniviridae / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D053585:Virus Attachment; D000511:alpha-Macroglobulins / Q000378:metabolism
chemical_list: D023181:Antimicrobial Cationic Peptides; D008565:Membrane Proteins; D015842:Serine Proteinase Inhibitors; D000511:alpha-Macroglobulins; C570319:crustinPm1 protein, Penaeus monodon
keywords: Biotinylation; Crustin; Hemocyte; Penaeus monodon; Yellow head virus