Denaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by guanidine hydrochloride; identification of inactive, partially unfolded, dimeric intermediates
dc.contributor.author | Plomer, J. Jeffrey | en_US |
dc.contributor.author | Gafni, Ari | en_US |
dc.date.accessioned | 2006-04-10T15:07:04Z | |
dc.date.available | 2006-04-10T15:07:04Z | |
dc.date.issued | 1992-08-21 | en_US |
dc.identifier.citation | Plomer, J. Jeffrey, Gafni, Ari (1992/08/21)."Denaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by guanidine hydrochloride; identification of inactive, partially unfolded, dimeric intermediates." Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology 1122(3): 234-242. <http://hdl.handle.net/2027.42/29896> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6T21-488GG9T-V/2/b7f38f9b9d2a4b75758f09cdd9fd8208 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/29896 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=1504085&dopt=citation | en_US |
dc.description.abstract | The denaturation of the dimeric enzyme glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by guanidine hydrochloride has been studied using enzymatic activity, intrinsic fluorescence, circular dichroism, and light scattering measurements. Equilibrium experiments at 25[deg]C revealed that between 0.9 and 1.2 m denaturant the enzyme underwent a conformational change, exposing tryptophan residues to solvent, with some loss of secondary structure and a complete loss of enzymatic activity but without dimer dissociation to subunits. This inactive, partially unfolded, dimeric intermediate was susceptible to slow aggregation, perhaps due to exposure of `sticky' hydrophobic stretches of the polypeptide chain. A second equilibrium transition, reflecting extensive unfolding and dimer dissociation, occurred only at denaturant concentrations above 1.4 M. Kinetics experiments demonstrated that in the denaturant concentration range of 1.7-1.9 M the fluorescence change occurred in two distinct steps. The first step involved a large, very rapid drop in fluorescence whose rate was strongly dependent on the denaturant concentration. This was followed by a small, relatively slow rise in the emission intensity, the rate of which was independent of denaturant concentration. Enzymatic activity was lost with a denaturant-concentration-dependent rate, which was approx. 3-times slower than the rate of the first step in fluorescence change. A denaturation mechanism incorporating several unfolding intermediates and which accounts for all the above results is presented and discussed. While the fully unfolded enzyme regained up to 55% of its original activity upon dilution of denaturant to a concentration that would be expected to support native enzyme, denaturation intermediates were able to reactivate only minimally and in fact were found to aggregate and precipitate out of solution. | en_US |
dc.format.extent | 881231 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | Denaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by guanidine hydrochloride; identification of inactive, partially unfolded, dimeric intermediates | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Materials Science and Engineering | en_US |
dc.subject.hlbsecondlevel | Chemistry | en_US |
dc.subject.hlbsecondlevel | Chemical Engineering | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.subject.hlbtoplevel | Engineering | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Institute of Gerontology and Department of Biological Chemistry, The University of Michigan, Ann Arbor, USA | en_US |
dc.contributor.affiliationum | Institute of Gerontology and Department of Biological Chemistry, The University of Michigan, Ann Arbor, USA | en_US |
dc.identifier.pmid | 1504085 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/29896/1/0000253.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1016/0167-4838(92)90398-W | en_US |
dc.identifier.source | Biochimica et Biophysica Acta | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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