title: Study of refolding of calf intestinal alkaline phosphatase.
abstract: Calf intestinal alkaline phosphatase (CIP) was denatured in 3.0 M guanidine hydrochloride for 2 h at 25 degrees C, before being diluted 20-fold with 0.1 M, pH 8.0, Tris-HCl buffer solution containing various effector molecules such as Mg2+, Zn2+, and nucleotide phosphate. The reactivation courses of the enzyme were investigated by the level of activity recovery, the recovery rate constant, and the relative standard deviation of the data. In the presence of effectors, the courses under reducing and nonreducing conditions of disulfide bonds of protein were compared. It was concluded that for CIP, Mg2+ is a more efficient inducer of reconstitution of the active site and appears to play a specific role. In addition, the present study discusses the differences in the refolding effectors between bacterial and mammalian enzymes.
authors.affiliation: Department of Chemistry, Capital University of Medical Science, Beijing 100054, People's Republic of China.
authors.forename: Xiao-Juan
authors.initials: XJ
authors.lastname: Tian
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Song
authors.forename: Shu-Lian
authors.initials: SL
authors.lastname: Yan
authors.forename: Ying-Xia
authors.initials: YX
authors.lastname: Zhang
authors.forename: Hai-Meng
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000469:Alkaline Phosphatase; D008274:Magnesium; D015032:Zinc