Document Detail

Redox cycling and kinetic analysis of single molecules of solution-phase nitrite reductase.
MedLine Citation:
PMID:  21969548     Owner:  NLM     Status:  MEDLINE    
Single-molecule measurements are a valuable tool for revealing details of enzyme mechanisms by enabling observation of unsynchronized behavior. However, this approach often requires immobilizing the enzyme on a substrate, a process which may alter enzyme behavior. We apply a microfluidic trapping device to allow, for the first time, prolonged solution-phase measurement of single enzymes in solution. Individual redox events are observed for single molecules of a blue nitrite reductase and are used to extract the microscopic kinetic parameters of the proposed catalytic cycle. Changes in parameters as a function of substrate concentration are consistent with a random sequential substrate binding mechanism.
Randall H Goldsmith; Leandro C Tabares; Dorota Kostrz; Christopher Dennison; Thijs J Aartsma; G W Canters; W E Moerner
Related Documents :
18638408 - Enzymatic, immunological and phylogenetic characterization of brucella suis urease.
19579248 - Characterisation of a recombinant nadp-dependent glycerol dehydrogenase from gluconobac...
2310748 - Characterization of ferrochelatase in kidney and erythroleukemia cells.
8147868 - Formylmethanofuran synthesis by formylmethanofuran dehydrogenase from methanobacterium ...
218748 - Direct evidence for the presence of a different converting enzyme in the hamster cheek ...
6487238 - Different respiratory activities of mitochondria isolated from the subendocardium and s...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2011-10-03
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  108     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2011 Oct 
Date Detail:
Created Date:  2011-10-19     Completed Date:  2011-12-30     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  17269-74     Citation Subset:  IM    
Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Alcaligenes / enzymology,  genetics
Amino Acid Substitution
Bacterial Proteins / chemistry,  genetics,  metabolism
Copper / chemistry
Fluorescence Resonance Energy Transfer
Microfluidic Analytical Techniques
Models, Molecular
Mutagenesis, Site-Directed
Nitrite Reductases / chemistry*,  genetics,  metabolism*
Protein Structure, Quaternary
Recombinant Proteins / chemistry,  genetics,  metabolism
Reg. No./Substance:
0/Bacterial Proteins; 0/Recombinant Proteins; 0/Solutions; 7440-50-8/Copper; EC 1.7.-/Nitrite Reductases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Evidence of denser MgSiO3 glass above 133 gigapascal (GPa) and implications for remnants of ultraden...
Next Document:  Full p53 transcriptional activation potential is dispensable for tumor suppression in diverse lineag...