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Acta Chimica Sinica >

2012 , Vol. 70 >Issue 04: 453 - 458

DOI: https://doi.org/10.6023/A1109161

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Investigation on MMP-12 Aggregation State in Urea Contained Solution by NMR and Fluorescence Spectroscopy

  • Zhang Linlin ,
  • Feng Yanye ,
  • Lan Wenxian ,
  • Cao Chunyang ,
  • Xu Diansheng ,
  • Yang Zhong
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  • a State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China;
    b Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China;
    c State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China

Received date: 2011-09-16

  Revised date: 2011-10-18

  Online published: 2012-03-07

Supported by

National Key Basic Research Program of China (No. 2009CB918600).

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Abstract

MMP-12 is a drug target for cancer therapy, but it’s over-expressed as inclusion bodies in E. coli To produce it in a large scale, in this paper, NMR, intrinsic tryptophan fluorescence, and ANS (8-anilinol-naphthalenesul fonic acid) fluorescence were employed to monitor its aggregation state to improve the refolding yield from inclusion bodies. The midpoint for the refolding (Cm≈4, mid-point of transition) of MMP-12 obtained from NMR experiments coincides with those from intrinsic tryptophan fluorescence and ANS fluorescence. These data suggest that the urea concentration from 4 mol/L to 3 mol/L is a key step in refolding process of MMP-12. Based on these results, we changed the refolding method from conventional step-wise dialysis to continuous dialysis, thus the refolding yield of MMP-12 from its inclusion bodies was doubled.

Key words： NMR; steady-state fluorescence; ANS; MMP-12; refolding

Cite this article

Zhang Linlin , Feng Yanye , Lan Wenxian , Cao Chunyang , Xu Diansheng , Yang Zhong . Investigation on MMP-12 Aggregation State in Urea Contained Solution by NMR and Fluorescence Spectroscopy[J]. Acta Chimica Sinica, 2012 , 70(04) : 453 -458 . DOI: 10.6023/A1109161

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