Molecular cloning and characterization of a lipopolysaccharide and beta-1,3-glucan binding protein from fleshy prawn (Fenneropenaeus chinensis).
Pattern recognition proteins (PRPs), such as lipopolysaccharide and beta-1,3-glucan binding protein (LGBP), have been identified in many animals and play a crucial role in invertebrate defense systems. In the current study, an LGBP gene was cloned from fleshy prawn (Fenneropenaeus chinensis, Fc-LGBP) utilizing homology cloning and RACE methods. The full cDNA of the Fc-LGBP gene in fleshy prawn was 1253bp in size with a deduced 366 amino acid protein that includes a glycosyl hydrolase domain. Northern blot and RT-PCR data suggested that Fc-LGBP mRNA was mostly synthesized in haemocytes and that the expression was down-regulated 24h post-injection of bacteria. In situ hybridization demonstrated that Fc-LGBP mRNA was only detected in haemocyte cytoplasm, with no detection in other tissues. The molecular weight of the purified recombinantly expressed Fc-LGBP was approximately 46kDa. Immunohistochemistry of haemocytes revealed that Fc-LGBP protein was localized on the membrane of most cells. Data from bacterial binding assays utilizing purified protein suggested that rFc-LGBP had strong binding activity to Gram-negative bacteria.
Du
Xin-Jun
XJ
School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
Zhao
Xiao-Fan
XF
Wang
Jin-Xing
JX
D000209:Acute-Phase Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D008070:Lipopolysaccharides / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D011509:Proteoglycans; D051192:Receptors, Pattern Recognition / Q000737:chemistry / Q000235:genetics; D047071:beta-Glucans / Q000378:metabolism*; D000099231:Lipopolysaccharide-Binding Protein
D000209:Acute-Phase Proteins; D002352:Carrier Proteins; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D051192:Receptors, Pattern Recognition; D047071:beta-Glucans; D000099231:Lipopolysaccharide-Binding Protein; C010770:polysaccharide-K