Document Detail


Micro-resonance Raman study of optically trapped Escherichia coli cells overexpressing human neuroglobin.
MedLine Citation:
PMID:  17867813     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe the possibility of using a microresonance Raman spectrometer combined with a microfluidic system and optical tweezers to study Escherichia coli (E. coli) overexpressing wild type (wt) neuroglobin (NGB) and its E7Leu mutant, respectively. NGB is a recently discovered heme protein and its function still is a matter of debate. So far, the protein has been studied in its purified form, and in vivo measurements on the single cell level could give more information. To study the feasibility of the combined techniques, the possibilities of the setup are investigated by taking spectra from single cells and clusters of cells. We find that the microresonance Raman technique enables studies of the wt NGB protein in a living cell under fluctuating aerobic and anaerobic conditions. E. coli cells overexpressing wt NGB are stable, and the reversible oxygenation-deoxygenation can be studied over a long period of time. Further, the experiment indicates the presence of an enzymatic system in the bacteria reducing the ferric form NGB. The study of E. coli cells overexpressing E7Leu NGB, on the other hand, gives insight into limiting factors of the setup, such as cell lysis, photoinduced chemistry, and protein concentrations.
Authors:
Kerstin Ramser; Wim Wenseleers; Sylvia Dewilde; Sabine Van Doorslaer; Luc Moens; Dag Hanstorp
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of biomedical optics     Volume:  12     ISSN:  1083-3668     ISO Abbreviation:  J Biomed Opt     Publication Date:    2007 Jul-Aug
Date Detail:
Created Date:  2007-09-17     Completed Date:  2007-11-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9605853     Medline TA:  J Biomed Opt     Country:  United States    
Other Details:
Languages:  eng     Pagination:  044009     Citation Subset:  IM    
Affiliation:
Göteborgs University, Department of Physics, SE-412 96 Göteborg, Sweden. kerstin.ramser@ltu.se
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MeSH Terms
Descriptor/Qualifier:
Escherichia coli / physiology*
Globins / genetics,  metabolism*
Humans
Microfluidic Analytical Techniques / methods*
Nerve Tissue Proteins / genetics,  metabolism*
Optical Tweezers*
Protein Engineering / methods
Spectrum Analysis, Raman / methods*
Chemical
Reg. No./Substance:
0/Nerve Tissue Proteins; 0/neuroglobin; 9004-22-2/Globins

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


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