Ion mobility-mass spectrometry of charge-reduced protein complexes reveals general trends in the collisional ejection of compact subunits

The Analyst ◽  
2015 ◽  
Vol 140 (20) ◽  
pp. 7020-7029 ◽  
Author(s):  
Russell E. Bornschein ◽  
Brandon T. Ruotolo
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Total Protein ◽  
Quaternary Structure ◽  
Protein Complexes ◽  
Complex Mixtures ◽  
Ion Mobility Mass Spectrometry ◽  
Cellular Functions ◽  
Multiprotein Complexes ◽  
Wide Range

Multiprotein complexes have been shown to play critical roles across a wide range of cellular functions, but most probes of protein quaternary structure are limited in their ability to analyze complex mixtures and polydisperse structures using small amounts of total protein.

Ion Mobility–Mass Spectrometry Reveals Long-Lived, Unfolded Intermediates in the Dissociation of Protein Complexes

2007 ◽  
Vol 46 (42) ◽  
pp. 8001-8004 ◽  
Author(s):  
Brandon T. Ruotolo ◽  
Suk-Joon Hyung ◽  
Paula M. Robinson ◽  
Kevin Giles ◽  
Robert H. Bateman ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Ion Mobility Mass Spectrometry

Ion Mobility Mass Spectrometry of Two Tetrameric Membrane Protein Complexes Reveals Compact Structures and Differences in Stability and Packing

2010 ◽  
Vol 132 (44) ◽  
pp. 15468-15470 ◽  
Author(s):  
Sheila C. Wang ◽  
Argyris Politis ◽  
Natalie Di Bartolo ◽  
Vassiliy N. Bavro ◽  
Stephen J. Tucker ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Ion Mobility Mass Spectrometry ◽  
Membrane Protein Complexes

scholarly journals Native Tandem and Ion Mobility Mass Spectrometry Highlight Structural and Modular Similarities in Clustered-Regularly-Interspaced Shot-Palindromic-Repeats (CRISPR)-associated Protein Complexes From Escherichia coli and Pseudomonas aeruginosa

2012 ◽  
Vol 11 (11) ◽  
pp. 1430-1441 ◽  
Author(s):  
Esther van Duijn ◽  
Ioana M. Barbu ◽  
Arjan Barendregt ◽  
Matthijs M. Jore ◽  
Blake Wiedenheft ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mass Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Acquired Resistance ◽  
Native Mass Spectrometry ◽  
Ion Mobility Mass Spectrometry ◽  
E Coli ◽  
Ribonucleoprotein Complexes ◽  
Specific Association

The CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) immune system of bacteria and archaea provides acquired resistance against viruses and plasmids, by a strategy analogous to RNA-interference. Key components of the defense system are ribonucleoprotein complexes, the composition of which appears highly variable in different CRISPR/Cas subtypes. Previous studies combined mass spectrometry, electron microscopy, and small angle x-ray scattering to demonstrate that the E. coli Cascade complex (405 kDa) and the P. aeruginosa Csy-complex (350 kDa) are similar in that they share a central spiral-shaped hexameric structure, flanked by associating proteins and one CRISPR RNA. Recently, a cryo-electron microscopy structure of Cascade revealed that the CRISPR RNA molecule resides in a groove of the hexameric backbone. For both complexes we here describe the use of native mass spectrometry in combination with ion mobility mass spectrometry to assign a stable core surrounded by more loosely associated modules. Via computational modeling subcomplex structures were proposed that relate to the experimental IMMS data. Despite the absence of obvious sequence homology between several subunits, detailed analysis of sub-complexes strongly suggests analogy between subunits of the two complexes. Probing the specific association of E. coli Cascade/crRNA to its complementary DNA target reveals a conformational change. All together these findings provide relevant new information about the potential assembly process of the two CRISPR-associated complexes.

Ion Mobility-Mass Spectrometry Reveals the Influence of Subunit Packing and Charge on the Dissociation of Multiprotein Complexes

2010 ◽  
Vol 82 (23) ◽  
pp. 9702-9710 ◽  
Author(s):  
Elisabetta Boeri Erba ◽  
Brandon T. Ruotolo ◽  
Daniel Barsky ◽  
Carol V. Robinson
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Ion Mobility Mass Spectrometry ◽  
Multiprotein Complexes

scholarly journals Integrating Ion Mobility Mass Spectrometry with Molecular Modelling to Determine the Architecture of Multiprotein Complexes

PLoS ONE ◽  
2010 ◽  
Vol 5 (8) ◽  
pp. e12080 ◽  
Author(s):  
Argyris Politis ◽  
Ah Young Park ◽  
Suk-Joon Hyung ◽  
Daniel Barsky ◽  
Brandon T. Ruotolo ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Molecular Modelling ◽  
Ion Mobility Mass Spectrometry ◽  
Multiprotein Complexes
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