Presentation Type

Poster

Abstract

Bartonella bacilliformis is a human bacterial pathogen and the etiological agent of Carrion’s Disease. B. bacilliformis is serum resistant, allowing it to survive in the human bloodstream and persist and replicate in erythrocytes. Human Factor H is a circulating protein in human blood that is part of the complement cascade of innate immune defense. Factor H binds to self-cells and prevents auto-immunity by complement fixation. A Far-Western blot followed by mass spectrometry analysis suggests that B. bacilliformis can bind to human Factor H protein. Here, we describe the protein BB1133, an outer membrane auto-transporter, and the Factor H—binding protein identified by mass spectrometry. By splitting the protein into two distinct domains, each domain is able to be expressed in E. coli via the Gateway cloning system and the pET-Dest42 vector. IPTG induction of the domains, followed by a Far-Western blot utilizing human Factor H as a probe, will indicate which domain of BB1133 is human Factor H—binding.

Category

Life Sciences

Available for download on Sunday, April 19, 2020

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Apr 28th, 3:00 PM Apr 28th, 4:00 PM

Characterization of a Bartonella Bacilliformis Human Factor H-Binding Protein

UC South Ballroom

Bartonella bacilliformis is a human bacterial pathogen and the etiological agent of Carrion’s Disease. B. bacilliformis is serum resistant, allowing it to survive in the human bloodstream and persist and replicate in erythrocytes. Human Factor H is a circulating protein in human blood that is part of the complement cascade of innate immune defense. Factor H binds to self-cells and prevents auto-immunity by complement fixation. A Far-Western blot followed by mass spectrometry analysis suggests that B. bacilliformis can bind to human Factor H protein. Here, we describe the protein BB1133, an outer membrane auto-transporter, and the Factor H—binding protein identified by mass spectrometry. By splitting the protein into two distinct domains, each domain is able to be expressed in E. coli via the Gateway cloning system and the pET-Dest42 vector. IPTG induction of the domains, followed by a Far-Western blot utilizing human Factor H as a probe, will indicate which domain of BB1133 is human Factor H—binding.