Identification of potential protein partners that bind to the variant surface glycoprotein in Trypanosoma equiperdum.
Trypanosoma equiperdum possesses a dense coat of a variant surface glycoprotein (VSG) that is used to evade the host immune response by a process known as antigenic variation. Soluble and membrane forms of the predominant VSG from the Venezuelan T. equiperdum TeAp-N/D1 strain (sVSG and mVSG, respectively) were purified to homogeneity; and antibodies against sVSG and mVSG were raised, isolated, and employed to produce anti-idiotypic antibodies that structurally mimic the VSG surface. Prospective VSG-binding partners were initially detected by far-Western blots, and then by immunoblots using the generated anti-idiotypic antibodies. Polypeptides of ~80 and 55 kDa were isolated when anti-idiotypic antibodies-Sepharose affinity matrixes were used as baits. Mass spectrometry sequencing yielded hits with various proteins from Trypanosoma brucei such as heat-shock protein 70, tryparedoxin peroxidase, VSG variants, expression site associated gene product 6, and two hypothetical proteins. In addition, a possible interaction with a protein homologous to the glutamic acid/alanine-rich protein from Trypanosoma congolense was also found. These results indicate that the corresponding orthologous gene products are candidates for VSG-interacting proteins in T. equiperdum.
Departamento de Biología Celular,Universidad Simón Bolívar,Caracas,Venezuela.
Liomary M
LM
Carrasquel
Departamento de Biología Celular,Universidad Simón Bolívar,Caracas,Venezuela.
José L
JL
Escalona
Parasitology Department,Liverpool School of Tropical Medicine,Liverpool,UK.
Alvaro
A
Acosta-Serrano
Department of Chemistry and Biochemistry,University of California San Diego,La Jolla, California 92093-0654,USA.
Yurong
Y
Guo
Departamento de Biología Celular,Universidad Simón Bolívar,Caracas,Venezuela.
José
http://orcid.org/0000-0002-8839-6745
J
Bubis
D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D014345:Trypanosoma / Q000378:metabolism*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism*
D015800:Protozoan Proteins; D014643:Variant Surface Glycoproteins, Trypanosoma

               Trypanosoma equiperdum
            ; anti-idiotypic antibodies; glycosylphosphatidylinositol-anchored proteins; protein–protein interactions; variant surface glycoprotein