Determining the stoichiometry and interactions of macromolecular assemblies from mass spectrometry
The growing number of applications to determine the stoichiometry, interactions and even subunit architecture of protein complexes from mass spectra suggests that some general guidelines can now be proposed. In this protocol, we describe the necessary steps required to maintain interactions between...
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| Veröffentlicht in: | Nature protocols 2007-03, Vol.2 (3), p.715-726 |
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| creator | Hernández, Helena Robinson, Carol V |
| description | The growing number of applications to determine the stoichiometry, interactions and even subunit architecture of protein complexes from mass spectra suggests that some general guidelines can now be proposed. In this protocol, we describe the necessary steps required to maintain interactions between subunits in the gas phase. We begin with the preparation of suitable solutions for electrospray (ES) and then consider the transmission of complexes through the various stages of the mass spectrometer until their detection. Subsequent steps are also described, including the dissociation of these complexes into multiple subcomplexes for generation of interaction networks. Throughout we highlight the critical experimental factors that determine success. Overall, we develop a generic protocol that can be carried out using commercially available ES mass spectrometers without extensive modification. |
| doi_str_mv | 10.1038/nprot.2007.73 |
| format | Article |
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| subjects | Analysis Analytical Chemistry Biological Techniques Biomedical and Life Sciences Computational Biology/Bioinformatics Investigations Life Sciences Mass spectra Mass spectrometry Methods Microarrays Multiprotein Complexes - chemistry Multiprotein Complexes - isolation & purification Organic Chemistry Proteins Proteomics Proteomics - methods Protocol Scientific imaging Spectrometers Spectrometry, Mass, Electrospray Ionization - instrumentation Spectrometry, Mass, Electrospray Ionization - methods Yeast |
| title | Determining the stoichiometry and interactions of macromolecular assemblies from mass spectrometry |
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