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Using flow cytometry to identify the orientations of P6 protein in Nontypable Haemophilus influenzae and Escherichia coli
Author(s) -
Snyder Joy,
Czup Katerina,
Sharma Sharad,
Pichichero Michael,
Michel Lea Vacca
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.581.2
Subject(s) - haemophilus influenzae , bacterial outer membrane , flow cytometry , escherichia coli , microbiology and biotechnology , biology , bacteria , extracellular , immunology , genetics , gene , antibiotics
Nontypable Haemophilus influenzae (NTHi) is a pathogenic bacteria and the cause of several illnesses, including pneumonia and otitis media. Thus, the development of a vaccine for NTHi would be highly beneficial. The outer membrane protein P6 is currently a top vaccine candidate for NTHi. Past studies have demonstrated that P6 can interact with molecules both inside AND outside of the cell, but its sequence and structure suggests that it does not cross the outer cellular membrane. Results from our flow cytometry experiments confirm that P6 is surface exposed and able to interact with extracellular antibodies. Other studies from our group have shown that P6 is also located inside of NTHi cells. Thus, we propose that P6 exhibits two orientations in the outer membrane of NTHi, a novel biological phenomenon only recently described for one other bacterial lipoprotein. We used flow cytometry to show that the P6 homolog, Pal, in Escherichia coli (E. coli) is also surface exposed, suggesting Pal also exhibits a dual orientation in the outer membrane of E. coli. This study was funded by the Rochester Institute of Technology, NIH NIDCD RO1 08671 (to MEP), and by an ASBMB UAN Undergraduate Research Award.