The identification and characterization of P5 and P2 colonization proteins in Haemophilus parasuis and the targeted binding of carcinoembryonic antigen (CEA)

Thumbnail Image
Date
2004-01-01
Authors
McVicker, Jerry
Major Professor
Advisor
Louisa B. Tabatabai
Committee Member
Journal Title
Journal ISSN
Volume Title
Publisher
Altmetrics
Authors
Research Projects
Organizational Units
Organizational Unit
Veterinary Microbiology and Preventive Medicine
Our faculty promote the understanding of causes of infectious disease in animals and the mechanisms by which diseases develop at the organismal, cellular and molecular levels. Veterinary microbiology also includes research on the interaction of pathogenic and symbiotic microbes with their hosts and the host response to infection.
Journal Issue
Is Version Of
Versions
Series
Abstract

The outer membrane protein P5 from Haemophilus influenzae has been shown to aid colonization in the human nasopharynx by binding to CD66 (CEA) on the surface of mucosal epithelial cells. The intent here was to determine if Haemophilus parasuis contained homologs of the P5 and P2 proteins and to characterize these proteins by SDS-PAGE, IEF, and CEA binding. Using outer membrane protein P5 primers designed from the H. influenzae database, we were able to detect a single PCR product in a H. parasuis field isolate. The PCR product was transformed into a competent E. coli strain and sequenced. DNA sequence analysis revealed homology to the H. influenzae P5 protein. N-terminal sequencing was used to confirm the presence of a 32 kDa P5 protein in both virulent and avirulent reference strains. Immunoblots were performed with H. parasuis reference strains representing all serovars using a P5 monoclonal antibody. A 48 kDa protein was identified in all but one virulent reference strain, whereas a 55 kDa protein was identified in all but one non-virulent reference strain. The 48 and 55 kDa proteins were subsequently identified as P2 by N-terminal sequencing. The outer membrane protein P5 from a Haemophilus parasuis field isolate (serovar 5) was fractionated by anion exchange and size-exclusion chromatography. The isolation of the pure P5 protein from the fraction was performed by elution from a polyacrylamide gel. Identification of the protein as P5 was determined by N-terminal sequencing. The P5 protein was further characterized by isoelectric focusing (IEF). Using a CEA immunoblotting method, it was determined that the purified P5 protein did not bind CEA. However, CEA did bind to a P2 protein in all type strain serovars. The molecular weight of the P2 protein varied between the virulent and avirulent serovars suggesting that it could potentially be used in discriminating between virulent and avirulent strains of H. parasuis.

Comments
Description
Keywords
Citation
Source
Copyright
Thu Jan 01 00:00:00 UTC 2004