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A comprehensive structure–function analysis shed a new light on molecular mechanism by which a novel smart copolymer, NY-3-1, assists protein refolding

Paper ID Volume ID Publish Year Pages File Format Full-Text
23858 43478 2012 7 PDF Available
Title
A comprehensive structure–function analysis shed a new light on molecular mechanism by which a novel smart copolymer, NY-3-1, assists protein refolding
Abstract

An in-depth understanding of molecular basis by which smart polymers assist protein refolding can lead us to develop a more effective polymer for protein refolding. In this report, to investigate structure–function relationship of pH-sensitive smart polymers, a series of poly(methylacrylic acid (MAc)-acrylic acid (AA))s with different MAc/AA ratios and molecular weights were synthesized and then their abilities in refolding of denatured lysozyme were compared by measuring the lytic activity of the refolded lysozyme. Based on our analysis, there were optimal MAc/AA ratio (44% MAc), Mw (1700 Da), and copolymer concentration (0.1%, w/v) at which the highest yield of protein refolding was achieved. Fluorescence, circular dichroism, and RP-HPLC analysis reported in this study demonstrated that the presence of P(MAc-AA)s in the refolding buffer significantly improved the refolding yield of denatured lysozyme without affecting the overall structure of the enzyme. Importantly, our bioseparation analysis, together with the analysis of zeta potential and particle size of the copolymer in refolding buffers with different copolymer concentrations, suggested that the polymer provided a negatively charged surface for an electrostatic interaction with the denatured lysozyme molecules and thereby minimized the hydrophobic-prone aggregation of unfolded proteins during the process of refolding.

► Novel smart copolymers, P(MAc-AA)s, were synthesized. ► An optimized P(MAc-AA), NY-3-1, significantly improved the refolding yield of denatured lysozyme. ► The ability of NY-3-1 in protein refolding was affected by its composition and molecular weight. ► The copolymer, NY-3-1, electrostatically engaged lysozyme during the process of refolding.

Keywords
P(MAc-AA)s; Lysozyme; Protein refolding; Molecular mechanism
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A comprehensive structure–function analysis shed a new light on molecular mechanism by which a novel smart copolymer, NY-3-1, assists protein refolding
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 160, Issues 3–4, 31 August 2012, Pages 169–175
Authors
, , , , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us