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Outer membrane proteome analysis of Indian strain of Pasteurella multocida serotype B:2 by MALDI-TOF/MS analysis.

Prasannavadhana A, Kumar S, Thomas P, Sarangi LN, Gupta SK, Priyadarshini A, Nagaleekar VK, Singh VP - ScientificWorldJournal (2014)

Bottom Line: The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis.Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition.Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2.

View Article: PubMed Central - PubMed

Affiliation: Division of Veterinary Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India.

ABSTRACT
Identification of outer membrane proteins (OMPs) is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS) in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.

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Related in: MedlinePlus

Immunodominant outer membrane proteins of Pasteurella multocida B:2 in field sera. Lane M: protein marker (250–10 kDa). Lane 1: immunodominant proteins in iron-limited condition. Lane 2: immunodominant protein in normal condition.
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fig4: Immunodominant outer membrane proteins of Pasteurella multocida B:2 in field sera. Lane M: protein marker (250–10 kDa). Lane 1: immunodominant proteins in iron-limited condition. Lane 2: immunodominant protein in normal condition.

Mentions: Western blot analysis showed the presence of immunodominant OMPs of P. multocida serotype B:2. It was found that the immunodominant protein bands obtained in the immunoblots differed with the type of antisera used as shown in Figures 2, 3, 4, and 5. A total of five immunodominant proteins, namely, 125, 70, 38, 36, and 23 kDa, were found using hyperimmune sera, four proteins 38, 36, 30, and 24 kDa using field sera, six proteins 38, 36, 35, 33, 32, and 23 kDa using vaccinated animal sera, and two proteins 38, 36 kDa using healthy animal sera. The details of the immunodominant bands are shown in Table 2.


Outer membrane proteome analysis of Indian strain of Pasteurella multocida serotype B:2 by MALDI-TOF/MS analysis.

Prasannavadhana A, Kumar S, Thomas P, Sarangi LN, Gupta SK, Priyadarshini A, Nagaleekar VK, Singh VP - ScientificWorldJournal (2014)

Immunodominant outer membrane proteins of Pasteurella multocida B:2 in field sera. Lane M: protein marker (250–10 kDa). Lane 1: immunodominant proteins in iron-limited condition. Lane 2: immunodominant protein in normal condition.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4283227&req=5

fig4: Immunodominant outer membrane proteins of Pasteurella multocida B:2 in field sera. Lane M: protein marker (250–10 kDa). Lane 1: immunodominant proteins in iron-limited condition. Lane 2: immunodominant protein in normal condition.
Mentions: Western blot analysis showed the presence of immunodominant OMPs of P. multocida serotype B:2. It was found that the immunodominant protein bands obtained in the immunoblots differed with the type of antisera used as shown in Figures 2, 3, 4, and 5. A total of five immunodominant proteins, namely, 125, 70, 38, 36, and 23 kDa, were found using hyperimmune sera, four proteins 38, 36, 30, and 24 kDa using field sera, six proteins 38, 36, 35, 33, 32, and 23 kDa using vaccinated animal sera, and two proteins 38, 36 kDa using healthy animal sera. The details of the immunodominant bands are shown in Table 2.

Bottom Line: The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis.Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition.Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2.

View Article: PubMed Central - PubMed

Affiliation: Division of Veterinary Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India.

ABSTRACT
Identification of outer membrane proteins (OMPs) is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS) in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.

Show MeSH
Related in: MedlinePlus