Mass spectrometry of ribosomes and ribosomal subunits.

Nanoflow electrospray ionization has been used to introduce intact Escherichia coli ribosomes into the ion source of a mass spectrometer. Mass spectra of remarkable quality result from a partial, but selective, dissociation of the particles within the mass spectrometer. Peaks in the spectra have been assigned to individual ribosomal proteins and to noncovalent complexes of up to five component proteins. The pattern of dissociation correlates strongly with predicted features of ribosomal protein-protein and protein-RNA interactions. The spectra allow the dynamics and state of folding of specific proteins to be investigated in the context of the intact ribosome. This study demonstrates a potentially general strategy to probe interactions within complex biological assemblies.

[1]  Richard D. Smith,et al.  Carbonic Anhydrase-Inhibitor Binding: From Solution to the Gas Phase , 1997 .

[2]  M. Rodnina,et al.  Hydrolysis of GTP by elongation factor G drives tRNA movement on the ribosome , 1997, Nature.

[3]  C. Spahn,et al.  Conserved Nucleotides of 23 S rRNA Located at the Ribosomal Peptidyltransferase Center* , 1996, The Journal of Biological Chemistry.

[4]  C. Dobson,et al.  Probing the Nature of Noncovalent Interactions by Mass Spectrometry. A Study of Protein−CoA Ligand Binding and Assembly , 1996 .

[5]  D. Smith,et al.  Mass spectrometric determination of isotopic exchange rates of amide hydrogens located on the surfaces of proteins. , 1996, Analytical chemistry.

[6]  I. Chernushevich,et al.  Probing the oligomeric structure of an enzyme by electrospray ionization time-of-flight mass spectrometry. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[7]  L Mayne,et al.  Mechanisms and uses of hydrogen exchange. , 1996, Current opinion in structural biology.

[8]  C. Dobson,et al.  Investigation of protein folding by mass spectrometry , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[9]  N. Volkmann,et al.  A milestone in ribosomal crystallography: the construction of preliminary electron density maps at intermediate resolution. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[10]  J. Frank,et al.  A model of protein synthesis based on cryo-electron microscopy of the E. coli ribosome , 1995, Nature.

[11]  C. Dobson,et al.  Conformation of GroEL-bound α-lactalbumin probed by mass spectrometry , 1994, Nature.

[12]  Richard D. Smith,et al.  Observation of the noncovalent quaternary associations of proteins by electrospray ionization mass spectrometry , 1994 .

[13]  C. Dobson,et al.  Detection of transient protein folding populations by mass spectrometry. , 1993, Science.

[14]  A. Bonincontro,et al.  Effects of magnesium ions on ribosomes: a fluorescence study. , 1993, Biochimica et biophysica acta.

[15]  John Skilling,et al.  Disentangling electrospray spectra with maximum entropy , 1992 .

[16]  H. Noller,et al.  Unusual resistance of peptidyl transferase to protein extraction procedures. , 1992, Science.

[17]  V. Katta,et al.  Conformational changes in proteins probed by hydrogen-exchange electrospray-ionization mass spectrometry. , 1991, Rapid communications in mass spectrometry : RCM.

[18]  F M Poulsen,et al.  A nuclear magnetic resonance study of the hydrogen-exchange behaviour of lysozyme in crystals and solution. , 1991, Journal of molecular biology.

[19]  M. Cannon.  Ribosomes and protein synthesis, a practical approach : Edited by G. Spedding; Oxford University Press; Oxford, 1990; xxiv + 318 pages; £22.50 , 1991 .

[20]  Brian T. Chait,et al.  Probing conformational changes in proteins by mass spectrometry , 1990 .

[21]  R. Brimacombe,et al.  A model for the spatial arrangement of the proteins in the large subunit of the Escherichia coli ribosome. , 1988, The EMBO journal.

[22]  J A Langer,et al.  A complete mapping of the proteins in the small ribosomal subunit of Escherichia coli. , 1987, Science.

[23]  J. Littlechild,et al.  The tertiary structure of salt-extracted ribosomal proteins from Escherichia coli as studied by proton magnetic resonance spectroscopy and limited proteolysis experiments. , 1987, Biochimica et biophysica acta.

[24]  A. Kerlavage,et al.  Reconstitution of Escherichia coli ribosomes containing puromycin-modified S14: functional effects of the photoaffinity labeling of a protein essential for tRNA binding. , 1986, Biochemistry.

[25]  J. Lake,et al.  Ribosomal proteins L7/L12 localized at a single region of the large subunit by immune electron microscopy. , 1978, Journal of molecular biology.

[26]  A. Liljas,et al.  The ribosomal protein L8 is a complex of L7/L12 and L10 , 1976, FEBS letters.

[27]  K. Aune,et al.  Molecular interactions between ribosomal proteins. Evidence for specificity of interaction between isolated proteins. , 1975, Biochemistry.

[28]  D. Draper,et al.  High resolution solution structure of ribosomal protein L11-C76, a helical protein with a flexible loop that becomes structured upon binding to RNA , 1997, Nature Structural Biology.

[29]  G. Siuzdak,et al.  Mass spectrometry and viral analysis. , 1996, Chemistry & biology.

[30]  M. Wilm,et al.  Analytical properties of the nanoelectrospray ion source. , 1996, Analytical chemistry.

[31]  J. Loo.  Observation of large subunit protein complexes by electrospray ionization mass spectrometry , 1995 .

[32]  A. Liljas.  Comparative biochemistry and biophysics of ribosomal proteins. , 1991, International review of cytology.

[33]  G. Spedding.  Ribosomes and protein synthesis : a practical approach , 1990 .