Polyethylene glycol enhanced refolding of the recombinant human tissue transglutaminase

Preparative Biochemistry & Biotechnology
A Ambrus, L Fésüs

Abstract

Tissue transglutaminase forms cross-links between lysine and glutamine side-chains of polypeptide chains in a Ca2+-dependent reaction; its structural basis is still not clarified. In this study, we demonstrate that the refolding of the human recombinant enzyme molecule to its catalytically active form from inclusion bodies needs the presence of a helper material with higher molecular mass, but only in the initiation phase. Ca2+ and nucleotides are ascribed as affector molecules also in the early phase of structural reconstitution. Two optimal concentrations of polyethylene glycol and a relatively long time scale for the evolution of the final structure were identified. The optimized refolding procedure is reported.

References

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Citations

Apr 2, 2002·Protein Expression and Purification·Qingli ShiArthur J L Cooper
Jan 9, 2004·Protein Expression and Purification·Steve M F G GilletJoelle N Pelletier
Jul 28, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Hamid R KalhorHossein Hosseinkhani

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Methods Mentioned

BETA
X-ray
Assay
electron microscopy
electrophoresis

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