Mass Spectrometry — a Key Technology in Proteom Research

Sickmann, Albert; Mreyen, Marcus; Meyer, Helmut E.

doi:10.1007/3-540-36459-5_6

Cited by 24 publications

(26 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Images of the membrane were captured by a CCD camera as a TIFF file and analyzed with image analysis software (GridGrinder, Corning Incorporated, Corning, NY) for spot intensity. The data were imported into a Microsoft Excel for graphing and statistical analysis Sample Preparation and Trypsin-mediated 18 O Labeling-Tryptic peptides generated from BPH and PCa cells were desalted using C-18 ZipTip microcolumns (Millipore, Billerica, MA), lyophilized to dryness, and resuspended in 0.1% TFA for nanoRPLC-MS/MS analysis. For the isotope labeling, protein extracts from an approximately equivalent number of microdissected cells were lyophilized and reconstituted separately in H 2 16 O (BPH) and H 2 18 O (PCa) each containing 20% methanol (v/v).…”

Section: Methodsmentioning

confidence: 99%

“…ac.uk/integr8) using SEQUEST (Thermo Electron). Peptides were searched using fully tryptic cleavage constraints and a dynamic 4.008-amu modification on the C terminus for the 18 O isotope-labeling analysis. For a peptide to be considered legitimately identified, it had to achieve stringent charge state and proteolytic cleavage-dependent cross-correlation (X corr ) scores of 1.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Proteomic Analysis of Formalin-fixed Prostate Cancer Tissue

Hood

Darfler

Guiel

et al. 2005

Molecular & Cellular Proteomics

263

275

View full text Add to dashboard Cite

Proteomic analysis of formalin-fixed paraffin-embedded (FFPE) tissue would enable retrospective biomarker investigations of this vast archive of pathologically characterized clinical samples that exist worldwide. These FFPE tissues are, however, refractory to proteomic investigations utilizing many state of the art methodologies largely due to the high level of covalently cross-linked proteins arising from formalin fixation. A novel tissue microdissection technique has been developed and combined with a method to extract soluble peptides directly from FFPE tissue for mass spectral analysis of prostate cancer (PCa) and benign prostate hyperplasia (BPH). Hundreds of proteins from PCa and BPH tissue were identified, including several known PCa markers such as prostate-specific antigen, prostatic acid phosphatase, and macrophage inhibitory cytokine-1. Quantitative proteomic profiling utilizing stable isotope labeling confirmed similar expression levels of prostate-specific antigen and prostatic acid phosphatase in BPH and PCa cells, whereas the expression of macrophage inhibitory cytokine-1 was found to be greater in PCa as compared with BPH cells.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Proteomic Analysis of Formalin-fixed Prostate Cancer Tissue

Hood

Darfler

Guiel

et al. 2005

Molecular & Cellular Proteomics

263

275

View full text Add to dashboard Cite

show abstract

“…The sequencing of the human genome, the development of gene arrays, and the availability of soft-ionization mass spectrometry techniques have led the way for highthroughput genomics and proteomics (6,7). Although the human genome comprises just over 25,000 genes, the human proteome, considering a multitude of posttranslational modifications, is composed of millions of different proteins.…”

Section: Supplementary Abstract Inflammation and Mass Spectrometry And Ementioning

confidence: 99%

Thematic review series: Systems Biology Approaches to Metabolic and Cardiovascular Disorders. Lipidomics: a global approach to lipid analysis in biological systems

Watson

2006

Journal of Lipid Research

414

246

View full text Add to dashboard Cite

Lipids are water-insoluble molecules that have a wide variety of functions within cells, including: 1) maintenance of electrochemical gradients; 2) subcellular partitioning; 3) first-and second-messenger cell signaling; 4) energy storage; and 5) protein trafficking and membrane anchoring. The physiological importance of lipids is illustrated by the numerous diseases to which lipid abnormalities contribute, including atherosclerosis, diabetes, obesity, and Alzheimer's disease. Lipidomics, a branch of metabolomics, is a systems-based study of all lipids, the molecules with which they interact, and their function within the cell. Recent advances in soft-ionization mass spectrometry, combined with established separation techniques, have allowed the rapid and sensitive detection of a variety of lipid species with minimal sample preparation. A ''lipid profile'' from a crude lipid extract is a mass spectrum of the composition and abundance of the lipids it contains, which can be used to monitor changes over time and in response to particular stimuli.Lipidomics, integrated with genomics, proteomics, and metabolomics, will contribute toward understanding how lipids function in a biological system and will provide a powerful tool for elucidating the mechanism of lipid-based disease, for biomarker screening, and for monitoring pharmacologic therapy.-Watson, A. D. Lipidomics: a global approach to lipid analysis in biological systems.

show abstract

“…More recently, mass spectrometry has emerged as a key technology for mapping protein phosphorylation sites, owing to its high sensitivity and speed of analysis (13)(14)(15)(16)(17). Mapping of phosphorylation sites within complex protein mixtures, however, is still a difficult task because of the often low stoichiometry of phosphorylation and signal suppression by unphosphorylated peptides.…”

mentioning

confidence: 99%

Phosphoproteome analysis of the human mitotic spindle

Nousiainen

Silljé

Sauer

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

303

260

View full text Add to dashboard Cite

During cell division, the mitotic spindle segregates the sister chromatids into two nascent cells, such that each daughter cell inherits one complete set of chromosomes. Errors in spindle formation can result in both chromosome missegregation and cytokinesis defects and hence lead to genomic instability. To ensure the correct function of the spindle, the activity and localization of spindle associated proteins has to be tightly regulated in time and space. Reversible phosphorylation has been shown to be one of the key regulatory mechanisms for the organization of the mitotic spindle. The relatively low number of identified in vivo phosphorylation sites of spindle components, however, has hampered functional analysis of regulatory spindle networks. A more complete inventory of the phosphorylation sites of spindle-associated proteins would therefore constitute an important advance. Here, we describe the mass spectrometry-based identification of in vivo phosphorylation sites from purified human mitotic spindles. In total, 736 phosphorylation sites were identified, of which 312 could be attributed to known spindle proteins. Among these are phosphorylation sites that were previously shown to be important for the regulation of spindle-associated proteins. Importantly, this data set also comprises 279 novel phosphorylation sites of known spindle proteins for future functional studies. This inventory of spindle phosphorylation sites should thus make an important contribution to a better understanding of the molecular mechanisms that regulate the formation, function, and integrity of the mitotic spindle. mass spectrometry ͉ mitosis ͉ molecular cell biology ͉ phosphorylation ͉ proteomics A t the transition from interphase to mitosis, the microtubule network undergoes a profound change that culminates in the formation of the spindle apparatus. The mitotic spindle then serves to segregate the chromosomes to opposite poles of the cell and to define the plane of cell division. A large number of proteins associate with this microtubule-based structure and regulate its dynamic formation and function (1, 2). Several spindle proteins, including XKCM1 and XMAP215 family members (3, 4), have been identified that either stabilize or destabilize microtubules by mediating the rapid changes between polymerization and depolymerization (5, 6). These proteins play an important role in spindle formation, as illustrated by the striking change in microtubule half-life from 5-10 min in interphase to less than 1 min in mitosis (7), which results in the short and unstable microtubules characteristic of mitosis (5, 6). Another important group of spindle proteins comprises motors of the kinesin and dynein families that are essential for mitotic progression (5,6,8,9). They push the spindle poles away from each other during early mitosis and play crucial roles in capturing chromosomes and positioning them at the metaphase plate. Subsequently, motors also contribute to central spindle formation and cytokinesis. In addition, numerous structural prote...

show abstract

Mass Spectrometry — a Key Technology in Proteom Research

Cited by 24 publications

References 54 publications

Proteomic Analysis of Formalin-fixed Prostate Cancer Tissue

Proteomic Analysis of Formalin-fixed Prostate Cancer Tissue

Thematic review series: Systems Biology Approaches to Metabolic and Cardiovascular Disorders. Lipidomics: a global approach to lipid analysis in biological systems

Phosphoproteome analysis of the human mitotic spindle

Contact Info

Product

Resources

About