NDLI: Stable Isotope Pulse-Chase Monitored by Quantitative Mass Spectrometry Applied to E. coli 30S Ribosome Assembly Kinetics

Content Provider	PubMed Central
Author	Bunner, Anne E. Williamson, James R.
Copyright Year	2009
Abstract	Stable isotope mass spectrometry has become a widespread tool in quantitative biology. Pulse-chase monitored by quantitative mass spectrometry (PC/QMS) is a recently developed stable isotope approach that provides a powerful means of studying the in vitro self-assembly kinetics of macromolecular complexes. This method has been applied to the E. coli 30S ribosomal subunit, but could be applied to any stable self-assembling complex that can be reconstituted from its component parts and purified from a mixture of components and complex. The binding rates of 18 out of the 20 ribosomal proteins have been measured at several temperatures using PC/QMS. Here, PC/QMS experiments on 30S ribosomal subunit assembly are described, and the potential application of the method to other complexes is discussed. A variation on the PC/QMS experiment is introduced that enables measurement of kinetic cooperativity between proteins. In addition, several related approaches to stable isotope labeling and quantitative mass spectrometry data analysis are compared and contrasted.
Related Links	http://dx.doi.org/10.1016/j.ymeth.2009.06.002
Ending Page	141
Page Count	6
Starting Page	136
File Format	PDF
ISSN	10462023
e-ISSN	10959130
Journal	Methods (San Diego, Calif.)
Issue Number	2
Volume Number	49
Language	English
Publisher Date	2009-10-01
Access Restriction	Open
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Molecular Biology

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