Derivatization of small biomolecules for optimized matrix‐assisted laser desorption/ionization mass spectrometry

Tholey, Andreas; Wittmann, Christoph; Kang, Min‐Jung; Bungert, Ditte; Hollemeyer, Klaus; Heinzle, Elmar

doi:10.1002/jms.355

Cited by 62 publications

(48 citation statements)

References 34 publications

(34 reference statements)

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“…Presumably, the PITC derivatives have diminished ESI ionization efficiency. This has indeed been observed to be true forÅMALDIÅionizationÅofÅPITCÅderivativesÅ [32].ÅThis problem might be solved by using modified Edman reagents designed for enhanced ESI ionization efficiency of the product phenylthiohydantoins used in EdmanÅ sequencingÅ [33]. Response relative to the most abundant (100) is given as the mean ϩ/Ϫ SD (n ϭ 8).…”

Section: Resultsmentioning

confidence: 99%

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

Onisko¹,

Dynin²,

Requena

et al. 2007

J. Am. Soc. Mass Spectrom.

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Sensitive quantitation of prions in biological samples is an extremely important and challenging analytical problem. Prions are the cause of several fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs). At this time, there are no methods to diagnose TSEs in live animals or to assure a prion-free blood supply for humans. Prions have been shown to be present in blood by transfusion experiments, but based on the amount of infectivity found in these types of experiments, the amount of misfolded prion protein in blood is estimated to be only 30 to 625 amol/mL. More sensitive detection of prions in brain would allow earlier detection of disease and assure a safer food supply. We studied quantitation of the prion protein by use of nanoscale liquid chromatography coupled to a tandem mass spectrometer using the multiple reaction monitoring mode of operation. We developed a method based on the detection of VVEQMCTTQYQK obtained by reduction, alkylation, and digestion with trypsin of the prion protein.

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Section: Resultsmentioning

confidence: 99%

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

Onisko¹,

Dynin²,

Requena

et al. 2007

J. Am. Soc. Mass Spectrom.

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“…Pyruvate was routinely derivatised with 2,4-dinitrophenylhydrazine, and measured photometrically at 450 nm; pyruvate was also determined enzymically with lactate dehydrogenase (Lamprecht & Heinz, 1984). Pyruvate was identified by MALDI-TOF MS after derivatization (Tholey et al, 2002). 3-Sulfopyruvate was determined as the azine formed by reaction with 2-(diphenylacetyl)indane-1,3-dione-1-hydrazone (Cunningham et al, 1998) after separation by HPLC with UV detection (400 nm).…”

Section: Methodsmentioning

confidence: 99%

Dissimilation of cysteate via 3-sulfolactate sulfo-lyase and a sulfate exporter in Paracoccus pantotrophus NKNCYSA

et al. 2005

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Paracoccus pantotrophus NKNCYSA utilizes (R)-cysteate (2-amino-3-sulfopropionate) as a sole source of carbon and energy for growth, with either nitrate or molecular oxygen as terminal electron acceptor, and the specific utilization rate of cysteate is about 2 mkat (kg protein) "1 .The initial degradative reaction is catalysed by an (R)-cysteate : 2-oxoglutarate aminotransferase, which yields 3-sulfopyruvate. The latter was reduced to 3-sulfolactate by an NAD-linked sulfolactate dehydrogenase [3?3 mkat (kg protein) "1 ]. The inducible desulfonation reaction was not detected initially in cell extracts. However, a strongly induced protein with subunits of 8 kDa (a) and 42 kDa (b) was found and purified. The corresponding genes had similarities to those encoding altronate dehydratases, which often require iron for activity. The purified enzyme could then be shown to convert 3-sulfolactate to sulfite and pyruvate and it was termed sulfolactate sulfo-lyase (Suy). A high level of sulfite dehydrogenase was also induced during growth with cysteate, and the organism excreted sulfate. A putative regulator, OrfR, was encoded upstream of suyAB on the reverse strand. Downstream of suyAB was suyZ, which was cotranscribed with suyB. The gene, an allele of tauZ, encoded a putative membrane protein with transmembrane helices (COG2855), and is a candidate to encode the sulfate exporter needed to maintain homeostasis during desulfonation. suyAB-like genes are widespread in sequenced genomes and environmental samples where, in contrast to the current annotation, several presumably encode the desulfonation of 3-sulfolactate, a component of bacterial spores.

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“…Moreover, the specific pentafluorobenzyl oxime derivatives produced by the carbonyl-reactive agent O-PFB enhance the detection sensitivity of carbonylcompounds (42,43). LC-MS/MS analysis of the different bacterial supernatant extracts identified hydroxyketones with a chain length from 10 to 17 carbon units (Fig.…”

Section: Prevalence Of Lqsa In L Pneumophila Strains and Genementioning

confidence: 99%

The Legionella Autoinducer Synthase LqsA Produces an α-Hydroxyketone Signaling Molecule

Spirig

Tiaden

Kiefer

et al. 2008

Journal of Biological Chemistry

107

114

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The opportunistic pathogen Legionella pneumophila replicates in human lung macrophages and in free-living amoebae. To accommodate the transfer between host cells, L. pneumophila switches from a replicative to a transmissive phase. L. pneumophila harbors a gene cluster homologous to the Vibrio cholerae cqsAS quorum sensing system, encoding a putative autoinducer synthase (lqsA) and a sensor kinase (lqsS), which flank a response regulator (lqsR). LqsR is an element of the L. pneumophila virulence regulatory network, which promotes pathogen-host cell interactions and inhibits entry into the replicative growth phase. Here, we show that lqsA functionally complements a V. cholerae cqsA autoinducer synthase deletion mutant and, upon expression in L. pneumophila or Escherichia coli, produces the diffusible signaling molecule LAI-1 (Legionella autoinducer-1). LAI-1 is distinct from CAI-1 (Cholerae autoinducer-1) and was identified as 3-hydroxypentadecan-4-one using liquid chromatography coupled to high resolution tandem mass spectrometry. The activity of both LqsA and CqsA was abolished upon mutation of a conserved lysine, and covalent binding of the cofactor pyridoxal 5-phosphate to this lysine was confirmed by mass spectrometry. Thus, LqsA and CqsA belong to a family of pyridoxal 5-phosphate-dependent autoinducer synthases, which produce the ␣-hydroxyketone signaling molecules LAI-1 and CAI-1.

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Derivatization of small biomolecules for optimized matrix‐assisted laser desorption/ionization mass spectrometry

Cited by 62 publications

References 34 publications

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

Dissimilation of cysteate via 3-sulfolactate sulfo-lyase and a sulfate exporter in Paracoccus pantotrophus NKNCYSA

The Legionella Autoinducer Synthase LqsA Produces an α-Hydroxyketone Signaling Molecule

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