Differential protein expression in ovaries of uninfected and Babesia-infected southern cattle ticks, Rhipicephalus (Boophilus) microplus.
We used gel electrophoresis and mass spectrometry to investigate differences in protein expression in ovarian tissues from Babesia bovis-infected and uninfected southern cattle tick, Rhipicephalus (Boophilus) microplus. Soluble and membrane proteins were extracted from ovaries of adult female ticks, and analyzed by isoelectric focusing (IEF) and one-dimensional or two-dimensional (2-D) gel electrophoresis. Protein patterns were analyzed for differences in expression between infected and uninfected ticks. 2-D separation of proteins revealed a number of proteins that appeared to be up- or down-regulated in response to infection with Babesia, in particular membrane/membrane-associated proteins and proteins in a low molecular mass range between 6 and 36kDa. A selection of differentially expressed proteins was subjected to analysis by capillary-HPLC-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). Among the ovarian proteins that were up-regulated in infected ticks were calreticulin, two myosin subunits, an endoplasmic reticulum protein, a peptidyl-prolyl cis-trans isomerase (PPIase), a cytochrome c oxidase subunit, a glutamine synthetase, and a family of Kunitz-type serine protease inhibitors. Among the down-regulated ovarian proteins were another PPIase, a hemoglobin subunit, and a lysozyme. This study is part of an ongoing effort to establish a proteome database that can be utilized to investigate specific proteins involved in successful pathogen transmission.
Rachinsky
Anna
A
USDA-ARS, Knipling-Bushland US Livestock Insects Research Laboratory, 2700 Fredericksburg Rd, Kerrville, TX 78028, USA. anna.rachinsky@ars.usda.gov
Guerrero
Felix D
FD
Scoles
Glen A
GA
D000595:Amino Acid Sequence; D000818:Animals; D016793:Babesia bovis / Q000502:physiology*; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005786:Gene Expression Regulation; D006790:Host-Parasite Interactions / Q000502:physiology; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010053:Ovary / Q000737:chemistry / Q000378:metabolism; D048494:Rhipicephalus / Q000378:metabolism* / Q000382:microbiology; D015854:Up-Regulation
D008565:Membrane Proteins