Abstract：
:We investigated the effect of homogenization strategy and protein precipitation on downstream protein quantitation using multiple reaction monitoring mass spectrometry (MRM-MS). Our objective was to develop a workflow capable of processing disparate tissue types with high throughput, minimal variability, and maximum purity. Similar abundances of endogenous proteins were measured in nine different mouse tissues regardless of the homogenization method used; however, protein precipitation had strong positive effects on several targets. The best throughput was achieved by lyophilizing tissues to dryness, followed by homogenization via bead-beating without sample buffer. Finally, the effect of tissue perfusion prior to dissection and collection was explored in 20 mouse tissues. MRM-MS showed decreased abundances of blood-related proteins in perfused tissues; however, complete removal was not achieved. Concentrations of nonblood proteins were largely unchanged, although significantly higher variances were observed for proteins from the perfused lung, indicating that perfusion may not be suitable for this organ. We present a simple yet effective tissue processing workflow consisting of harvest of fresh nonperfused tissue, novel lyophilization and homogenization by bead-beating, and protein precipitation. This workflow can be applied to a range of mouse tissues with the advantages of simplicity, minimal manual manipulation of samples, use of commonly available equipment, and high sample quality.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Michaud SA,Pětrošová H,Jackson AM,McGuire JC,Sinclair NJ,Ganguly M,Flenniken AM,Nutter LMJ,McKerlie C,Schibli D,Smith D,Borchers CHdoi
10.1021/acs.jproteome.0c00399subject
Has Abstractpub_date
2021-01-01 00:00:00pages
305-316issue
1eissn
1535-3893issn
1535-3907journal_volume
20pub_type
杂志文章abstract：:Leukocytes play an important role in the progression of disease and leukocyte-derived proteins are associated with the pathogenesis of the disease. Leukocyte activation causes production of inflammatory mediators, over-expression of cell surface adhesion molecules, and an increase in migration and infiltration, phagoc...
journal_title：Journal of proteome research
pub_type： 杂志文章,评审
doi：10.1021/pr0499601
更新日期：2004-09-01 00:00:00
abstract：:Human milk oligosaccharides (HMOs) perform a number of functions including serving as prebiotics to stimulate the growth of beneficial intestinal bacteria, as receptor analogues to inhibit binding of pathogens, and as substances that promote postnatal brain development. There is further evidence that HMOs participate ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100362f
更新日期：2010-08-06 00:00:00
abstract：:Human plasma and serum proteins are subject to intrinsic proteolytic degradation both during and after blood collection. By monitoring peptides, we investigated the stability of plasma and serum samples and the effects of anticoagulants and protease inhibitors on the plasma samples. Serum and plasma were subjected to ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060550h
更新日期：2007-05-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b01019
更新日期：2017-12-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060157b
更新日期：2006-09-01 00:00:00
abstract：:A novel linear ion trap (LIT) mass spectrometer with dual matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) ionization sources has been built in the MALDI-LIT-ESI configuration. The design features two independent ion source/ion optical channels connected to opposite ends of a singl...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060514i
更新日期：2007-02-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00343
更新日期：2019-10-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00118
更新日期：2020-07-02 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900383g
更新日期：2009-10-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00699
更新日期：2018-01-05 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800507x
更新日期：2009-01-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8003024
更新日期：2008-10-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900918h
更新日期：2010-03-05 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00327
更新日期：2015-11-06 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章,评审
doi：10.1021/pr300125t
更新日期：2012-05-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00419
更新日期：2015-08-07 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800619h
更新日期：2009-03-01 00:00:00
abstract：:As a result of the complexity and dynamic range of the cellular proteome, including mutual interactions and interactions with other molecules, focused proteomic approaches are important to study subsets of physiologically important proteins. In one such approach, a small molecule or part of a protein is immobilized on...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060278b
更新日期：2006-09-01 00:00:00
abstract：:The low selectivity of Kv1 peptide inhibitors for specific isoforms makes them poor candidates for the development of theraputics. Using combined approaches, we showed that the Kv1 turret is the critical determinant for ADWX-1 peptide inhibitor selectivity of Kv1.3 over Kv1.1. Mutation of Kv1.1 turret residues to matc...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800494a
更新日期：2008-11-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8006478
更新日期：2009-01-01 00:00:00
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pub_type： 杂志文章
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journal_title：Journal of proteome research
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更新日期：2006-06-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2007-05-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2014-11-07 00:00:00
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journal_title：Journal of proteome research
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doi：10.1021/pr200039p
更新日期：2011-07-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00615
更新日期：2020-04-03 00:00:00
abstract：:Proteomics of human milk has been used to identify the comprehensive cargo of proteins involved in immune and cellular function. Very little is known about the effects of gestational diabetes mellitus (GDM) on lactation and breast milk components. The objective of the current study was to examine the effect of GDM on ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500818d
更新日期：2015-01-02 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2012-02-03 00:00:00
abstract：:Energy imbalance is a primary cause of obesity. While the classical approach to attenuate weight gain includes an increase in energy expenditure through exercise, dietary manipulation such as the inclusion of dairy products has also been proven effective. In the present study, we explored the potential mechanisms by w...
journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2019-11-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr050122w
更新日期：2005-07-01 00:00:00
