Journal article
Glyceraldehyde-3-Phosphate Dehydrogenase Is a Surface-Associated, Fibronectin-Binding Protein of Trichomonas vaginalis
Infection and immunity, Vol.77(7), pp.2703-2711
07/2009
Handle:
https://hdl.handle.net/2376/110595
PMCID: PMC2708568
PMID: 19380472
Abstract
Trichomonas vaginalis
colonizes the urogenital tract of humans and causes trichomonosis, the most prevalent nonviral sexually transmitted disease. We have shown an association of
T. vaginalis
with basement membrane extracellular matrix components, a property which we hypothesize is important for colonization and persistence. In this study, we identify a fibronectin (FN)-binding protein of
T. vaginalis
. A monoclonal antibody (MAb) from a library of hybridomas that inhibited the binding of
T. vaginalis
organisms to immobilized FN was identified. The MAb (called ws1) recognized a 39-kDa protein and was used to screen a cDNA expression library of
T. vaginalis
. A 1,086-bp reactive cDNA clone that encoded a protein of 362 amino acids with identity to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was obtained. The
gapdh
gene was cloned, and recombinant GAPDH (rGAPDH) was expressed in
Escherichia coli
cells. Natural GAPDH and rGAPDH bound to immobilized FN and to plasminogen and collagen but not to laminin. MAb ws1 inhibited binding to FN. GAPDH was detected on the surface of trichomonads and was upregulated in synthesis and surface expression by iron. Higher levels of binding to FN were seen for organisms grown in iron-replete medium than for organisms grown in iron-depleted medium. In addition, decreased synthesis of GAPDH by antisense transfection of
T. vaginalis
gave lower levels of organisms bound to FN and had no adverse effect on growth kinetics. Finally, GAPDH did not associate with immortalized vaginal epithelial cells (VECs), and neither GAPDH nor MAb ws1 inhibited the adherence of trichomonads to VECs. These results indicate that GAPDH is a surface-associated protein of
T. vaginalis
with alternative functions.
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Details
- Title
- Glyceraldehyde-3-Phosphate Dehydrogenase Is a Surface-Associated, Fibronectin-Binding Protein of Trichomonas vaginalis
- Creators
- A Lama - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164A Kucknoor - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164V Mundodi - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164J. F Alderete - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164
- Publication Details
- Infection and immunity, Vol.77(7), pp.2703-2711
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- American Society for Microbiology (ASM)
- Identifiers
- 99900547225101842
- Language
- English
- Resource Type
- Journal article