title: Polyethylene glycol enhanced refolding of the recombinant human tissue transglutaminase.
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.
authors.affiliation: University of Debrecen, Department of Biochemistry and Molecular Biology, Hungary.
authors.forename: A
authors.initials: A
authors.lastname: Ambrus
authors.forename: L
authors.initials: L
authors.lastname: Fésüs
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006801:Humans; D009711:Nucleotides / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry; D011503:Transglutaminases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D009711:Nucleotides; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D004492:Edetic Acid; D011503:Transglutaminases; D002118:Calcium