Identification of GST Interacted Proteins under PRSV Infected Papaya Using Affinity Purification–mass Spectrometry

Abstract

Glutathione S-transferases (GSTs) are multifunctional proteins involved in stress metabolism, which play major roles in biotic and abiotic stress responses. GSTs have found in all organisms which is a major phase II detoxification enzymes found in the cytosol. GSTs regulate peroxidase and isomerase activities, they protect cells against H(2)O(2)-induced cell death. Papaya rigsport virus (PRSV) is one of main biotic agent that cause damage in papaya. The disease symptoms are mosaic, chrolosis, ring spot and stunt, all the symptoms are the early stage of cell death. The main point of this research to investigate protein interaction of the PRSV interacted plant proteins and GSTs recombinant protein using the classical Affinity-purification-mass spectrometry (AP-MS) approache. The GST protein was heterologous expressed in E.coli system and the pull down assay was applied to expore the protein interacted complexs after that the protein complexs were indentified by LC-MS/MS. CTC1, Protein CCA1 isoform X1, Tetratricopeptide repeat (TPR)-like superfamily protein, PHD finger protein ALFIN-LIKE 9 and Fructose-bisphosphate aldolase-lysine N-methyltransferase proteins were identified. The interacted five proteins were predicted the protein network by STITCH program, the result show they associated with oxidative stress response mechanism. However, this is the basic intensive information that could develop to manufacture PRSV resistance variety in the future.