Paper Information

Title: 

CHARACTERISATION OF AN ANTI-INFLAMMATORY PROTEIN (VOX2) FOR EVALUATING OF ITS ACTIVITIES IN MOLECULAR LEVEL

Type: SPEECH
Author(s): REZAEI S.A.,MILBURN C.,BLACKBOURN D.J.
 
 
 
Name of Seminar: IRANIAN CONGRESS OF PHYSIOLOGY AND PHARMACOLOGY
Type of Seminar:  CONGRESS
Sponsor:  PHYSIOLOGY AND PHARMACOLOGY SOCIETY, MASHHAD UNIVERSITY OF MEDICAL SCIENCE
Date:  2007Volume 18
 
 
Abstract: 

We have previously shown that KSHV vOX2 protein has anti-inflammatory activities. To reveal the structure of vOX2 and test its activities in a molecular level, different constructs of ORFK14 were cloned in frame with a gene encoding the C-terminal domain of the human Fcγ1. The resulting fusion proteins, vOX2:Fc and K14:Fc, were expressed from stable CHO cells and were then purified from with affinity chromatography and gel filtration and its identity confirmed by mass spectrometry. Purified vOX2:Fc was then cleaved and Fc tag removed using the Factor Xa Cleavage Capture Kit and the resulted bands were identified by MS. Moreover the N-linked oligosaccharides from the purified vOX2 fragment of the Fc fusion protein were released with PNGase F. These approaches revealed that vOX2 is an unstable protein which following deglycosylation of extracellular domain, the 50 kDa diffuse band of vOX2, shifted to a sharp band of 25 kDa. Therefore, vOX2 is a highly glycosylated protein of 50 kDa. In addition, we have shown that methionine 78 in the published sequence is the start codon and thransmembrane vOX2 is a protein of 271 amino acids (accession: NC_009333.1 <http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=NC_009333.1> protein ID: YP_001129432 <http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=protein&val=139472886>) and not a protein of 348 as Rusoo et al suggested. Taken together, this anti-inflammatory protein, vOX2, is a highly glaycosylated unstable transmembrane protein, containing an RGD motif and an Ig-Like domain. The cytoplasmic region of the molecule has no sequence identity with known signalling molecules, nor docking sites for adapter molecules Therefore, The extracellular part of the vOX2 might represent the biologically important region of vOX2 through binding to a cellular receptor.

 
Keyword(s): 
 
Yearly Visit 30   tarjomyar
 
Latest on Blog
Enter SID Blog