title: Folding and association of beta-Galactosidase.
abstract: beta-D-Galactosidase from Escherichia coli is one of the largest tetrameric enzymes known at present. Although its physiological importance, the regulation of its synthesis, its enzymatic properties and its structure are well established, little is known about the stability and the folding pathway of this enzyme. Here we show that the overall folding mechanism of chemically denatured beta-galactosidase consists of three stages: (i) formation of elements of secondary structure; (ii) collapse to subdomains and structured monomers; (iii) association to the native quaternary structure via dimeric intermediates. The first rate-limiting step is the association of structured monomers to form dimers in a bi-molecular reaction, with a rate constant of 4.3x10(3) M-1 s-1 at 20 degreesC. The second rate-limiting uni-molecular folding step leads to dimers which are competent for further association, with a rate constant of 0.5x10(-3) s-1 at 20 degreesC. Tetramers form from these dimers in a fast reaction. By determining a similar mechanism for alpha-complementation of beta-galactosidase fragments it could be confirmed that beta-galactosidase follows a consecutive bi-uni-molecular mechanism of folding and association.
authors.affiliation: Institut für Biophysik und physikalische Biochemie, Universität Regensburg, Universitätstr. 31, Regensburg, 93040, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Nichtl
authors.forename: J
authors.initials: J
authors.lastname: Buchner
authors.forename: R
authors.initials: R
authors.lastname: Jaenicke
authors.forename: R
authors.initials: R
authors.lastname: Rudolph
authors.forename: T
authors.initials: T
authors.lastname: Scheibel
mesh_terms: D019281:Dimerization; D004789:Enzyme Activation; D004795:Enzyme Stability; D005816:Genetic Complementation Test; D007700:Kinetics; D008956:Models, Chemical; D009154:Mutation; D017510:Protein Folding; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D001616:beta-Galactosidase