Peer Reviewed: Microwave-Enhanced Chemistry

Richter, Robert; Link, Dirk D.; Kingston, H. M.

doi:10.1021/ac0123781

Cited by 91 publications

(39 citation statements)

References 15 publications

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“…Microwave-assisted chemistry is a relatively new field. It has been shown to accelerate synthetic organic reactions [98,99]. Application of microwave technology to faster analysis of proteins/peptides is an emerging field that is gaining more attention.…”

Section: Microwave-assisted Methods For Proteins/peptidesmentioning

confidence: 99%

LC/MS Analysis of Proteins and Peptides in Drug Discovery

Chen¹,

Liu²,

Pramanik³

2006

HPLC for Pharmaceutical Scientists

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Section: Microwave-assisted Methods For Proteins/peptidesmentioning

confidence: 99%

LC/MS Analysis of Proteins and Peptides in Drug Discovery

Chen¹,

Liu²,

Pramanik³

2006

HPLC for Pharmaceutical Scientists

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“…Furthermore, a protein sam-ple does not need to be completely dissolved in TFA for MAAH-merely suspending the protein sample in TFA is sufficient for acid hydrolysis [21,37]. However, in the traditional microwave experiment using a household microwave oven [13,14,38], a sample vial containing a low volume of solution (40 L) only absorbs a small fraction of the microwave energy. The solution temperatures in the vials placed at different locations might be different, resulting in different rates of hydrolysis.…”

Section: New Setup For Maahmentioning

confidence: 99%

Reproducible microwave-assisted acid hydrolysis of proteins using a household microwave oven and its combination with LC-ESI MS/MS for mapping protein sequences and modifications

Wang

2010

J. Am. Soc. Mass Spectrom.

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A new set-up for microwave-assisted acid hydrolysis (MAAH) with high efficiency and reproducibility to degrade proteins into peptides for mass spectrometry analysis is described. It is based on the use of an inexpensive domestic microwave oven and can be used for low volume protein solution digestion. This set-up has been combined with liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI QTOF MS) for mapping protein sequences and characterizing phosphoproteins. It is demonstrated that for bovine serum albumin (BSA), with a molecular mass of about 67,000 Da, 1292 peptides (669 unique sequences) can be detected from a 2 g hydrolysate generated by trifluoroacetic acid (TFA) MAAH. These peptides cover the entire protein sequence, allowing the identification of an amino acid substitution in a natural variant of BSA. It is shown that for a simple phosphoprotein containing one phosphoform, ␤-casein, direct analysis of the hydrolysate generates a comprehensive peptide map that can be used to identify all five known phosphorylation sites. For characterizing a complex phosphoprotein consisting of different phosphoforms with varying numbers of phosphate groups and/or phosphorylation sites, such as bovine ␣ S1 -casein, immobilized metal-ion affinity chromatography (IMAC) is used to enrich the phosphopeptides from the hydrolysate, followed by LC-ESI MS analysis. The MS/MS data generated from the initial hydrolysate and the phosphopeptide-enriched fraction, in combination with MS analysis of the intact protein sample, allow us to reveal the presence of three different phosphoforms of bovine ␣ S1 -casein and assign the phosphorylation sites to each phosphoform with high confidence. (J Am Soc Mass Spectrom 2010, 21, 1573-1587) © 2010 American Society for Mass Spectrometry P rotein sequence mapping is commonly used to study post-translational modifications of a protein or amino acid substitutions from point mutations in the genome. Ideally, the entire amino acid sequence of a protein should be mapped to pinpoint where a modified amino acid or a substitution is located. Mass spectrometry (MS) has become an indispensable tool for protein sequence mapping [1,2]. This is usually done by using a top-down or a bottom-up proteomic approach [1][2][3][4][5][6][7][8][9][10][11][12]. In the top-down method, a protein ion is dissociated in a tandem mass spectrometer (MS/MS) and the fragment ions generated are interpreted to generate a stretch of amino acid sequence information. Sequence coverage by this method is dependent on the nature of the protein, ranging from a few residues to a full sequence [12]. In general, full sequence information is difficult to obtain for a protein with molecular mass of Ͼ20,000 Da [12]. The bottom-up approach is a robust method for protein identification based on sequencing by MS/MS one or more peptides generated by chemical or enzymatic degradation of a protein. For a protein digest, such as that produced by trypsin digestion, one or a few peptides are sequenced, r...

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“…The most important is the dipole rotation as a function of the oscillate electromagnetic field. At 2450 MHz, the dipoles align and randomise five billion times a second (Richter et al 2001). Also, the ionic conduction mechanism takes place, and the ionic species migrate in the direction of the electric field.…”

Section: Microwave-assisted Sample Preparationmentioning

confidence: 99%

Section: Microwave-assisted Sample Preparationmentioning

confidence: 99%

“…Such techniques require samples that are homogeneous at the molecular level in the sample solution (Richter et al 2001). Microwave chemistry is a fast, efficient and reproducible way to sample preparation for environmental purposes (Richter et al 2001). Also, there are clear advantages over more traditional technologies such as better recoveries of volatile compounds, lower contamination levels, minimum volumes of reagents, and a better working environment (Agazzi and Pirola 2000).…”

Section: Microwave-assisted Sample Preparationmentioning

confidence: 99%

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