Mass spectrometry peptides

U. Kertscher, M. Beyermann, E. Krause, J. Furkert, H. Berger, M. Bienert, B. Mehlis, The Degradation of Corticotropin-Releasing Factor by Enzymes of the Rat Brain Studied by Liquid Chromatography-Mass Spectrometry , Peptides 1998, 19, 649-658. 

Humeny A, Kislinger T, Becker CM, Pischetsrieder M. Qualitative detennination of specific protein glycation products by matrix-assisted laser desorption/ionization mass spectrometry Peptide mapping. J Agric Food Client 2002 50(7) 2153—2160. 

The methods for each study are divided into the initial protein separation step, a second separation step if applicable, the type of mass analysis, and the software used for peptide identification. ID = one dimensional polyacrylamide gel electrophoresis, 2D = two dimensional polyacrylamide gel electrophoresis, MS = mass spectrometry (peptide mass fingerprinting), MS/MS = tandem mass spectrometry, MALDI-TOF = matrix assisted laser desorption/ionization-time of flight, MS FIT = software from Protein Prospector, http //prospector.ucsf edu/, ESI = electrospray ionization, Q-TOF = quadrupole-time of flight, PPSS2 =Protana s Proteomic Software Suite (ProtanaEngineering, Odense, Denmark), Mascot = Matiix Science, http //www.matrixscience.com/, TOF-TOF = MALDI plus TOF tandem mass spectrometry, RP-HPLC = reverse phase high performance liquid chromatography, Q-IT = quadrupole ion trap, LIT = linear ion trap. Bioworks = Thermo Electron Corporation. 

MALDI-TOF Mass Spectrometry. Peptide/detergent mixtures as well as purified peptides were analyzed using a Vestec LaserTec BenchTop II system (PerSeptive Biosystems). Peptides were mixed with alpha-cyano-4-hydroxycinnamic acid as previously described (4). 

Figure 2. Peptide maps (A-C) and MALDI-TOF mass spectra (D-F) of PVDF-bound transferrin (53 pmol) digested with trypsin in the presence of 50 pi of 1% RTX-100/10% acetonitrile/100 mM Tris, pH 8.0 (A,D), 1% octylglucopyranoside/10% acetonitrile/100 mM Tris, pH 8.0 (B,E), and 1% decylglucopyranoside/10% acetonitrile/100 mM Tris, pH 8.0 (CJF) as described in Materials and Methods. Ninety percent of the digestion was analyzed by HPLC ( 29 pmol based on Table I) and 0.5% ( 150 fmol) was used for MALDI-TOF mass spectrometry. Peptides 1 and 2 in A-C were amino terminally sequenced (Table II) and analyzed by MALDI-TOF mass spectrometry (Figure 3).

K McMellop, W Davidson, G Hansen, D Freeman, N Pallai. The characterization of crude products from solid-phase peptide synthesis by v-HPLC/fast atom bombardment mass spectrometry. Peptide research 4 40-46, 1991. 

Medzihradszky, K.F., H. Leffler, M.A. Baldwin and A. Burlinghame. Protein identification by in-gel digestion, high-performance liquid chromatography and mass spectrometry peptide analysis by complementary ionization techniques. J. Am. Soc. Mass Spectrom. 12 215-221, 2001. 

McMahon, G. Mass spectrometry peptides and proteins. Encyclopedia of Analytical Science (2nd ed), Eds P Worsfold, A Townshend and C Poole. ISBN 012764100-9. Elsevier, 501-509 (2005). 

Although this technique was introduced in early 1990s, it was the introduction of a MALDI-TOF instrument capable of 50 ppm mass accuracy that made PMF a routine procedure. In MALDI-TOF mass spectrometry, peptides appear as singly charged species in the mass spectrum (Figure 4.4-1). Unlike an electrospray (ESI) mass spectrum, which displays multiply charged species, the MALDI-TOF spectrum is simple to interpret. PMF can also be used to identify proteins in ESI spectra, but it is seldom used because the peptide masses would need to be decon-voluted for each search. 

Traditional methods to generate peptide maps involve fractionation of complex mixtures of peptides in a protein digest either with one-dimensional SDS-PAGE or RP-HPLC [28,29]. The mass spectrometry peptide-mapping protocol, in principle, is similar to these techniques, but it provides an added dimension of structure-specific data (i.e., the molecular mass). MALDI-MS [30,31], ESl-MS [32], LC/ESI-MS [33], and CE/ESI-MS [34] have currently replaced the traditional biochemical approaches. MALDI allows the direct analysis of unfractionated protein digests. The commonly used matrices are sinapinic acid, a-cyano-4-hydroxy cinnamic acid (a-CHCA), and 2,5-dihydroxybenzoic acid (DHB). 

Oven/iew Waters, Sediments, and Soils. Ion-Selective Electrodes Water Applications. Isotope Dilution Analysis. Liquid Chromatography Size-Exclusion Liquid Chromatography-Mass Spectrometry Mass Spectrometry Peptides and Proteins. Voltammetry Overview. 

See also Electrophoresis Nucleic Acids. Mass Spectrometry Peptides and Proteins. 

See alsa Chromatography Multidimensional Techniques. Environmental Analysis. Extraction Solid-Phase Extraction. Food and Nutritional Analysis Sample Preparation Contaminants Pesticide Residues. Forensic Sciences Drug Screening in Sport Illicit Drugs. Herbicides. Liquid Chromatography Instrumentation Clinical Applications Food Applications. Mass Spectrometry Peptides and Proteins. Pesticides. Pharmaceutical Analysis Sample Preparation. Proteomics. Sample Handling Automated Sample Preparation. Water Analysis Organic Compounds. 

See also Capillary Electrophoresis Overview. Chir-optical Analysis. Liquid Chromatography Column Technology Mobile Phase Selection Reversed Phase Instrumentation Amino Acids. Mass Spectrometry Peptides and Proteins. Nuclear Magnetic Resonance Spectroscopy Techniques Nuclear Overhauser Effect. Proteins Traditional Methods of Sequence Determination Foods. 

Fig. 5. Representative peptide mass fingerprint for the 33.2 and 66.6 kDa proteins resulting from an in-gel digestion of the 66.6 kDa gel band obtained by MALDI-TOF mass spectrometry. Peptides marked with an asterisk result from autodigestion products of recombinant trypsin. Peptides labeled with a diamond were selected for MS/MS experiments carried out on a MALDI-QIT/RTOF and a nano-ESI-QIT mass spectrometer.

Organometallics Studied Using Mass Spectrometry Peptides and Proteins Studied Using Mass Spectrometry SIFT Applications in Mass David Smith and Patrik Spectrometry Stereochemistry Studied Using Mass Spectrometry... 

See also Chemical Structure Information from Mass Spectrometry Chromatography-MS, Methods Fast Atom Bombardment Ionization in Mass Spectrometry Fragmentation In Mass Spectrometry Hyphenated Techniques, Applications of in Mass Spectrometry Medical Applications of Mass Spectrometry MS-MS and MS Nucieic Acids and Nucieotides Studied Using Mass Spectrometry Peptides and Proteins Studied Using Mass Spectrometry Quadrupoies, Use of in Mass Spectrometry Sector Mass Spectrometers Surface induced Dissociation in Mass Spectrometry Time of Fiight Mass Spectrometers. 

See also Biochemical Applications of Mass MS-MS and MS Organometallics Studied Using Spectrometry Fragmentation in Mass Spectrometry Mass Spectrometry Peptides and Proteins Studied Ion Energetics in Mass Spectrometry Ionization The- Using Mass Spectrometry Plasma Desorption loniza-ory IR Spectroscopy Sample Preparation Methods tion in Mass Spectrometry Spectroscopy of Ions. 

Mass spectrometry peptide identification and quantification software (e.g., MaxQuant [5] or Proteome Discoverer Software, Thermo Scientific). 

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