TY - JOUR
T1 - Identifying components of protein complexes in C. elegans using co-immunoprecipitation and mass spectrometry
AU - Moresco, James J.
AU - Carvalho, Paulo C.
AU - Yates, John R.
N1 - Funding Information:
PCC is supported by Convenio CAPES / Fiocruz (Brazil) 30/2006, and the Center for Technological Development in Health of the Oswaldo Cruz Foundation from Rio de Janeiro, Brazil .
Funding Information:
JJM and JRY are supported by grant nos. DK074798 and AG027463 from the National Institutes of Health .
PY - 2010/10/10
Y1 - 2010/10/10
N2 - Mass spectrometry-based proteomics is rapidly becoming an essential tool for biologists. One of the most common applications is identifying the components of protein complexes isolated by co-immunoprecipitation. In this review, we discuss the co-immunoprecipitation, mass spectrometry and data analysis techniques that have been used successfully to define protein complexes in C. elegans research. In this discussion, two strategies emerged. One approach is to use stringent biochemical purification methods and attempt to identify a small number of complex components with a high degree of certainty based on MS data. A second approach is to use less stringent purification and identification parameters, and ultimately test a longer list of potential binding partners in biological validation assays. This should provide a useful guide for biologists planning proteomic experiments.
AB - Mass spectrometry-based proteomics is rapidly becoming an essential tool for biologists. One of the most common applications is identifying the components of protein complexes isolated by co-immunoprecipitation. In this review, we discuss the co-immunoprecipitation, mass spectrometry and data analysis techniques that have been used successfully to define protein complexes in C. elegans research. In this discussion, two strategies emerged. One approach is to use stringent biochemical purification methods and attempt to identify a small number of complex components with a high degree of certainty based on MS data. A second approach is to use less stringent purification and identification parameters, and ultimately test a longer list of potential binding partners in biological validation assays. This should provide a useful guide for biologists planning proteomic experiments.
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U2 - 10.1016/j.jprot.2010.05.008
DO - 10.1016/j.jprot.2010.05.008
M3 - Review article
C2 - 20546956
AN - SCOPUS:77957254319
SN - 1874-3919
VL - 73
SP - 2198
EP - 2204
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 11
ER -