Analysis of Detergent-Insoluble and Whole Cell Lysate Fractions of Resting Neutrophils Using High-Resolution Mass Spectrometry
Contribuinte(s) |
UNIVERSIDADE DE SÃO PAULO |
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Data(s) |
19/10/2012
19/10/2012
2010
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Resumo |
Neutrophilic granulocytes play a major role in the initiation and resolution of the inflammatory response, and demonstrate significant transcriptional and translational activity. Although much was known about neutrophils prior to the introduction of proteomics, the use of MS-based methodologies has provided an unprecedented tool to confirm and extend previous findings. In the present study, we performed a Gel-LC-MS/MS analysis of neutrophil detergent insoluble and whole cell lysate fractions of resting neutrophils. We achieved a set of identifications through the use of high-resolution mass spectrometry and validation of its data. We identified a total of 1249 proteins with a wide range of intensities from both detergent-insoluble and whole cell lysate fractions, allowing a mapping of proteins such as those involved in intracellular transport (Rab and Sec family proteins) and cell signaling (S100 proteins). These results represent the most comprehensive proteomic characterization of resting human neutrophils to date, and provide important information relevant for further studies of the immune system in health and disease. The methods applied here can be employed to help us understand how neutrophils respond to various physiologic and pathophysiologic conditions and could be extended to protein quantitation after cell activation. Norwegian Research Council[175141] Norwegian Research Council[423725] Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) |
Identificador |
JOURNAL OF PROTEOME RESEARCH, v.9, n.4, p.2030-2036, 2010 1535-3893 http://producao.usp.br/handle/BDPI/23814 10.1021/pr1000253 |
Idioma(s) |
eng |
Publicador |
AMER CHEMICAL SOC |
Relação |
Journal of Proteome Research |
Direitos |
restrictedAccess Copyright AMER CHEMICAL SOC |
Palavras-Chave | #proteomic profiling #neutrophils #cell fractionation #whole cell lysate #high resolution mass spectrometry #shotgun proteomics #GLYCATION END-PRODUCTS #PROTEOMIC ANALYSIS #SUBCELLULAR-LOCALIZATION #PROTEINS #RECEPTOR #S100C #INHIBITION #ACCURACY #GRANULES #ADHESION #Biochemical Research Methods |
Tipo |
article original article publishedVersion |