Studies of interaction of tumor suppressor p53 with apo-MT using surface plasmon resonance.
The interaction of tumor suppressor p53 with apo-metallothionein (apo-MT) has been carried out using a flow injection-surface plasmon resonance (FI-SPR) instrument. MT was first tethered onto the carboxymethylated dextran film. Via incorporation of glycine-HCl (pH 2) to remove the sequestrated metal ions inherent in MT molecules, a more extended and open structure of apo-MT was formed. Substantial SPR angle shift corresponding to the interaction of wild-type p53 with apo-MT was observed. The interaction was originated from the binding between the free sulfhydryl groups of apo-MT and Zn(2+) of p53 with the binding constant of 1.4 x 10(8) M(-1). The specific binding of p53 to consensus double-stranded DNA was hindered after metal chelation from p53 by apo-MT. Furthermore, inhibition of the interaction between p53 and apo-MT imposed by p53/DNA complex was observed. The fluorescence measurements also revealed the binding of p53 to apo-MT, being consistent with the SPR results. Thus, SPR could potentially serve as an attractive technique for monitoring p53 conformational change and transcriptional activity regulated by the MT/apo-MT couple.
School of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan, 410083, People's Republic of China.
Ning
N
Xia
Lin
L
Liu
Xinyao
X
Yi
Jianxiu
J
Wang
D000818:Animals; D015374:Biosensing Techniques; D004247:DNA / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000378:metabolism; D008668:Metallothionein / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D020349:Surface Plasmon Resonance / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000378:metabolism*
D016159:Tumor Suppressor Protein p53; D004247:DNA; D008668:Metallothionein