title: Mycobacteriophage cell binding proteins for the capture of mycobacteria.
abstract: Slow growingMycobacteriumaviumsubsp.paratuberculosis(MAP) causes a deadly condition in cattle known as Johne's disease where asymptomatic carriers are the major source of disease transmission. MAP was also shown to be associated with chronic Crohn's disease in humans.Mycobacterium smegmatisis a model mycobacterium that can cause opportunistic infections in a number of human tissues and, rarely, a respiratory disease. Currently, there are no rapid, culture-independent, reliable and inexpensive tests for the diagnostics of MAP orM. smegmatisinfections. Bacteriophages are viruses producing a number of proteins that effectively and specifically recognize the cell envelopes of their bacterial hosts. We demonstrate that the mycobacterial phage L5 minor tail protein Gp6 and lysin Gp10 are useful tools for the rapid capture of mycobacteria. Immobilized Gp10 was able to bind both MAP andM. smegmatiscells whereas Gp6 wasM. smegmatisspecific. Neither of the 2 proteins was able to captureE. coli, salmonella, campylobacter orMycobacterium marinumcells. Gp6 was detected previously as a component of the phage particle and shows no homology to proteins with known function. Therefore, electrospray ionization mass spectrometry was used to determine whether recombinant Gp6 could bind to a number of chemically synthesized fragments of mycobacterial surface glycans. These findings demonstrate that mycobacteriophage proteins could be used as a pathogen capturing platform that can potentially improve the effectiveness of existing diagnostic methods.
authors.affiliation: Department of Biological Sciences; University of Alberta , Edmonton, AB Canada ; Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada.
authors.forename: Denis
authors.initials: D
authors.lastname: Arutyunov
authors.affiliation: Department of Electrical and Computer Engineering; University of Alberta ; Edmonton, AB Canada.
authors.forename: Upasana
authors.initials: U
authors.lastname: Singh
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: Amr
authors.initials: A
authors.lastname: El-Hawiet
authors.affiliation: Department of Biological Sciences; University of Alberta , Edmonton, AB Canada ; Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada.
authors.forename: Henrique Dos Santos
authors.initials: HD
authors.lastname: Seckler
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: Sanaz
authors.initials: S
authors.lastname: Nikjah
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: Maju
authors.initials: M
authors.lastname: Joe
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: Yu
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: Todd L
authors.initials: TL
authors.lastname: Lowary
authors.affiliation: Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada ; Department of Chemistry; University of Alberta ; Edmonton, AB Canada.
authors.forename: John S
authors.initials: JS
authors.lastname: Klassen
authors.affiliation: Department of Electrical and Computer Engineering; University of Alberta ; Edmonton, AB Canada.
authors.forename: Stephane
authors.initials: S
authors.lastname: Evoy
authors.affiliation: Department of Biological Sciences; University of Alberta , Edmonton, AB Canada ; Alberta Glycomics Center; University of Alberta ; Edmonton, AB Canada.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Szymanski
keywords: Crohn's disease; Johne's disease; bacteriophage; lysin; mycobacterium; receptor binding protein