NDLI: Detection and selective dissociation of intact ribosomes in a mass spectrometer.

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Detection and selective dissociation of intact ribosomes in a mass spectrometer.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Benjamin, D. R. Rostom, A. A. Juenemann, R. Dobson, C. M. Robinson, C. V. Hartl, F. U. Fucini, P. Nierhaus, K. H.
Description	Author Affiliation: Rostom AA ( Oxford Centre for Molecular Sciences, New Chemistry Laboratory, University of Oxford, South Parks Road, Oxford, OX1 3QT, United Kingdom.);
Abstract	Intact Escherichia coli ribosomes have been projected into the gas phase of a mass spectrometer by means of nanoflow electrospray techniques. Species with mass/charge ratios in excess of 20,000 were detected at the level of individual ions by using time-of-flight analysis. Once in the gas phase the stability of intact ribosomes was investigated and found to increase as a result of cross-linking ribosomal proteins to the rRNA. By lowering the Mg(2+) concentration in solutions containing ribosomes the particles were found to dissociate into 30S and 50S subunits. The resolution of the charge states in the spectrum of the 30S subunit enabled its mass to be determined as 852,187 +/- 3,918 Da, a value within 0.6% of that calculated from the individual proteins and the 16S RNA. Further dissociation into smaller macromolecular complexes and then individual proteins could be induced by subjecting the particles to increasingly energetic gas phase collisions. The ease with which proteins dissociated from the intact species was found to be related to their known interactions in the ribosome particle. The results show that emerging mass spectrometric techniques can be used to characterize a fully functional biological assembly as well as its isolated components.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	97
Language	English
Publisher	National Academy of Sciences
Publisher Date	2000-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Escherichia Coli Ultrastructure Ribosomes Cross-Linking Reagents Magnesium Pharmacology Mass Spectrometry Molecular Weight RNA, Ribosomal Chemistry Ribosomal Proteins Drug Effects Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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