Identification and purification of resveratrol targeting proteins using immobilized resveratrol affinity chromatography.
The phytochemical resveratrol (trans-3,4',5-trihydroxystilbene) is a naturally occurring polyphenol with a plethora of health-beneficial properties, including a preventive role in cancer. We surmise that resveratrol may exert its diverse biological effects by interacting with specific target proteins, denoted RTPs. To test this possibility, resveratrol was immobilized on epoxy-activated agarose forming a resveratrol affinity column (RAC), which was used to detect and isolate RTPs. Distinct RTPs can be resolved on RAC by fractionation with increasing NaCl, followed by 1mM ATP, and finally, with 1-2mM resveratrol. A 22-kDa polypeptide, RTP-22, eluted with resveratrol was identified by MALDI-TOF MS and cloning/expression in Escherichia coli, as dihydronicotinamide riboside quinone reductase 2 (NQO2). The utility of RAC was additionally explored with extracts derived from different staging prostate cancer cells. NQO2 was most abundant in CWR22Rv1, a model for prostate cancer transition from androgen-dependent to the hormone-refractory state, but was marginally expressed in JCA-1 cells as representing more advanced stage prostate cancer. These results provide evidence for the existence of distinctive RTPs in mammalian cells and that RAC is a facile approach to identify and purify RTPs.
Wang
Zhirong
Z
Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA.
Hsieh
Tze-chen
TC
Zhang
Zhongtao
Z
Ma
Yuliang
Y
Wu
Joseph M
JM
D000327:Adsorption; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D016503:Drug Delivery Systems / Q000379:methods; D004353:Drug Evaluation, Preclinical; D006801:Humans; D008297:Male; D009363:Neoplasm Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D000077185:Resveratrol; D013267:Stilbenes / Q000032:analysis* / Q000737:chemistry / Q000627:therapeutic use
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D013267:Stilbenes; D000077185:Resveratrol