Method for identification and quantitative analysis of protein lysine methylation using matrix-assisted laser desorption/ionization — time-of-flight mass spectrometry and amino acid analysis

Yan, Jun X.; Sanchez, Jean‐Charles; Binz, Pierre‐Alain; Williams, Keith L.; Hochstrasser, Denis F.

doi:10.1002/(sici)1522-2683(19990101)20:4/5<749::aid-elps749>3.0.co;2-v

Cited by 25 publications

(10 citation statements)

References 21 publications

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“…In addition, many proteins were predicted to be phosphorylated (e.g., spots 208, 231, 234). Methylation [18] and phosphorylation (e.g., [19,20]) are two of the most commonly found modifications in a cell. Additional predicted modifications included sulfatation (e.g., spot 56), palmytoilation (e.g., spot 27), farnesylation (e.g., spot 99) and b-methylthiolation (e.g., spot 27).…”

Section: Multiple Spots For the Same Proteinsmentioning

confidence: 99%

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

Porubleva

Velden²,

Kothari³

et al. 2001

Electrophoresis

140

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As a first step in establishing a proteome database for maize, we have embarked on the identification of the leaf proteins resolved on two-dimensional (2-D) gels. We detected nearly 900 spots on the gels with a pH 4-7 gradient and over 200 spots on the gels with a pH 6-11 gradient when the proteins were visualized with colloidal Coomassie blue. Peptide mass fingerprints for 300 protein spots were obtained with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer and 149 protein spots were identified using the protein databases. We also searched the pdbEST databases to identify the leaf proteins and verified 66% of the protein spots that had been identified using the protein databases. Sixty-seven additional protein spots were identified from expressed sequence tags (ESTs). Many abundant leaf proteins are present in multiple spots. Functions of over 50% of the abundant leaf proteins are either unknown or hypothetical. Our results show that EST databases in conjunction with peptide mass fingerprints can be used for identifying proteins from organisms with incomplete genome sequence information.

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Section: Multiple Spots For the Same Proteinsmentioning

confidence: 99%

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

Porubleva

Velden²,

Kothari³

et al. 2001

Electrophoresis

140

View full text Add to dashboard Cite

show abstract

“…Total amino acid analysis (AAA) uses complete protein hydrolysis, derivatization, and chromatography to isolate and quantify each constituent amino acid [4,5]. The amount of each amino acid is then summed to obtain the protein mass with present sensitivities to the 0.1-1 pmol range [6]. Exchange of natural sulfur for radioactive 35 S (t 1/2 = 87.2 days) in methionine residues of isolated proteins [7] quantifies multi-femtomole quantities with laborious normalizations that include AAA and knowing the methionine content of the quantified protein [8].…”

Section: Quantitation Of Proteins In Gelsmentioning

confidence: 99%

Attomole quantitation of protein separations with accelerator mass spectrometry

et al. 2001

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Quantification of specific proteins depends on separation by chromatography or electrophoresis followed by chemical detection schemes such as staining and fluorophore adhesion. Chemical exchange of short-lived isotopes, particularly sulfur, is also prevalent despite the inconveniences of counting radioactivity. Physical methods based on isotopic and elemental analyses offer highly sensitive protein quantitation that has linear response over wide dynamic ranges and is independent of protein conformation. Accelerator mass spectrometry quantifies long-lived isotopes such as 14C to subattomole sensitivity. We quantified protein interactions with small molecules such as toxins, vitamins, and natural biochemicals at precisions of 1-5%. Micro-proton-induced X-ray emission quantifies elemental abundances in separated metalloprotein samples to nanogram amounts and is capable of quantifying phopsphorylated loci in gels. Accelerator-based quantitation is a possible tool for quantifying the genome translation into proteome.

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“…The number of studies, however, where post-translational modifications on proteins have been elucidated directly from gels, are few. These include, for example, site-specific assignment of protein phosphorylation [14±16] and protein glycosylation [17,18], characterization of N-linked glycans [19], and methylation [20].…”

Section: Introductionmentioning

confidence: 99%

Characterization of differently processed forms of enolase 2 fromSaccharomyces cerevisiae by two-dimensional gel electrophoresis and mass spectrometry

Larsen

Fey

et al. 2001

Electrophoresis

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Two-dimensional gel electrophoresis, bioinformatics, and mass spectrometry are key analysis tools in proteome analysis. The further characterization of post-translational modifications in gel-separated proteins relies fully on data obtained by mass spectrometric analysis. In this study, stress-induced changes in protein expression in Saccharomyces serevisiae were investigated. A total of eleven spots on a silver-stained two-dimensional (2-D) gel were identified by matrix-assisted laser desorption/ionization (MALDI) peptide mass mapping to represent C and/or N-terminal processed forms of enolase 2. The processing sites were determined by MALDI peptide mass mapping using a variety of proteolytic enzymes, by optimizing the sample preparation procedure and by specific labeling of all C-termini derived from in-gel digestion using a buffer containing 16O:18O (1:1). Out of eleven processed forms of enolase 2, six were fully characterized and the approximate processing sites identified for the remaining five.

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Method for identification and quantitative analysis of protein lysine methylation using matrix-assisted laser desorption/ionization — time-of-flight mass spectrometry and amino acid analysis

Cited by 25 publications

References 21 publications

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

Attomole quantitation of protein separations with accelerator mass spectrometry

Characterization of differently processed forms of enolase 2 fromSaccharomyces cerevisiae by two-dimensional gel electrophoresis and mass spectrometry

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