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https://www.arca.fiocruz.br/handle/icict/12130
SURFACEOME OF LEPTOSPIRA SPP.
Author
Affilliation
Australian Bacterial Pathogenesis Program and Victorian Bioinformatics Consortium. / Monash University. Department of Microbiology. Australia.
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California / David Geffen School of Medicine at UCLA. Department of Medicine. Los Angeles, California
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Weil Medical College of Cornell University. Division of International Medicine and Infectious Diseases. New York, New York
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California / David Geffen School of Medicine at UCLA. Department of Medicine. Los Angeles, California
Australian Bacterial Pathogenesis Program and Victorian Bioinformatics Consortium. / Monash University. Department of Microbiology. Australia.
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California / David Geffen School of Medicine at UCLA. Department of Medicine. Los Angeles, California
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Weil Medical College of Cornell University. Division of International Medicine and Infectious Diseases. New York, New York
Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California / David Geffen School of Medicine at UCLA. Department of Medicine. Los Angeles, California
Australian Bacterial Pathogenesis Program and Victorian Bioinformatics Consortium. / Monash University. Department of Microbiology. Australia.
Abstract
The identification of the subset of outer membrane proteins exposed on the surface of a bacterial cell (the
surfaceome) is critical to understanding the interactions of bacteria with their environments and greatly
narrows the search for protective antigens of extracellular pathogens. The surfaceome of Leptospira was
investigated by biotin labeling of viable leptospires, affinity capture of the biotinylated proteins, two-dimensional
gel electrophoresis, and mass spectrometry (MS). The leptospiral surfaceome was found to be predominantly
made up of a small number of already characterized proteins, being in order of relative abundance on
the cell surface: LipL32 > LipL21 > LipL41. Of these proteins, only LipL32 had not been previously identified
as surface exposed. LipL32 surface exposure was subsequently verified by three independent approaches:
surface immunofluorescence, whole-cell enzyme-linked immunosorbent assay (ELISA), and immunoelectron
microscopy. Three other proteins, Q8F8Q0 (a putative transmembrane outer membrane protein) and two
proteins of 20 kDa and 55 kDa that could not be identified by MS, one of which demonstrated a high degree
of labeling potentially representing an additional, as-yet-uncharacterized, surface-exposed protein. Minor
labeling of p31LipL45, GroEL, and FlaB1 was also observed. Expression of the surfaceome constituents remained
unchanged under a range of conditions investigated, including temperature and the presence of serum
or urine. Immunization of mice with affinity-captured surface components stimulated the production of
antibodies that bound surface proteins from heterologous leptospiral strains. The surfaceomics approach is
particularly amenable to protein expression profiling using small amounts of sample (<107 cells) offering the
potential to analyze bacterial surface expression during infection.
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