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Mass spectrometry-assisted protease substrate screening

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Mass spectrometry-assisted protease substrate screening. / Schlüter, Hartmut; Rykl, Jana; Thiemann, Joachim; Kurzawski, Sandra; Gobom, Johan; Tepel, Martin; Zidek, Walter; Linscheid, Michael.

in: ANAL CHEM, Jahrgang 79, Nr. 3, 01.02.2007, S. 1251-5.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Schlüter, H, Rykl, J, Thiemann, J, Kurzawski, S, Gobom, J, Tepel, M, Zidek, W & Linscheid, M 2007, 'Mass spectrometry-assisted protease substrate screening', ANAL CHEM, Jg. 79, Nr. 3, S. 1251-5. https://doi.org/10.1021/ac061482l

APA

Schlüter, H., Rykl, J., Thiemann, J., Kurzawski, S., Gobom, J., Tepel, M., Zidek, W., & Linscheid, M. (2007). Mass spectrometry-assisted protease substrate screening. ANAL CHEM, 79(3), 1251-5. https://doi.org/10.1021/ac061482l

Vancouver

Schlüter H, Rykl J, Thiemann J, Kurzawski S, Gobom J, Tepel M et al. Mass spectrometry-assisted protease substrate screening. ANAL CHEM. 2007 Feb 1;79(3):1251-5. https://doi.org/10.1021/ac061482l

Bibtex

@article{7519f8badebe47b5b4df7aea16c8998a,
title = "Mass spectrometry-assisted protease substrate screening",
abstract = "Since sequencing of the human genome was completed, more than 500 genes have been annotated as proteases. Exploring the physiological role of each protease requires the identification of their natural substrates. However, the endogenous substrates of many of the human proteases are as yet unknown. Here we describe a new assay that addresses this problem. The assay, which easily can be automated, is based on the incubation of immobilized protein fractions, which may contain the natural substrate, with a defined protease. After concentrating the proteolytically released peptides by reversed-phase chromatography they are analyzed by tandem mass spectrometry and the substrates identified by database searching. The proof of principle in this study is demonstrated by incubating immobilized human plasma proteins with thrombin and by identifying by tandem mass spectrometry the fibrinopeptides, released by the action of thrombin from their natural substrate fibrinogen, in the reaction mixture.",
keywords = "Blood Proteins, Fibrinogen, Humans, Peptide Hydrolases, Peptides, Substrate Specificity, Tandem Mass Spectrometry, Thrombin, Journal Article, Research Support, Non-U.S. Gov't",
author = "Hartmut Schl{\"u}ter and Jana Rykl and Joachim Thiemann and Sandra Kurzawski and Johan Gobom and Martin Tepel and Walter Zidek and Michael Linscheid",
year = "2007",
month = feb,
day = "1",
doi = "10.1021/ac061482l",
language = "English",
volume = "79",
pages = "1251--5",
journal = "ANAL CHEM",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "3",

}

RIS

TY - JOUR

T1 - Mass spectrometry-assisted protease substrate screening

AU - Schlüter, Hartmut

AU - Rykl, Jana

AU - Thiemann, Joachim

AU - Kurzawski, Sandra

AU - Gobom, Johan

AU - Tepel, Martin

AU - Zidek, Walter

AU - Linscheid, Michael

PY - 2007/2/1

Y1 - 2007/2/1

N2 - Since sequencing of the human genome was completed, more than 500 genes have been annotated as proteases. Exploring the physiological role of each protease requires the identification of their natural substrates. However, the endogenous substrates of many of the human proteases are as yet unknown. Here we describe a new assay that addresses this problem. The assay, which easily can be automated, is based on the incubation of immobilized protein fractions, which may contain the natural substrate, with a defined protease. After concentrating the proteolytically released peptides by reversed-phase chromatography they are analyzed by tandem mass spectrometry and the substrates identified by database searching. The proof of principle in this study is demonstrated by incubating immobilized human plasma proteins with thrombin and by identifying by tandem mass spectrometry the fibrinopeptides, released by the action of thrombin from their natural substrate fibrinogen, in the reaction mixture.

AB - Since sequencing of the human genome was completed, more than 500 genes have been annotated as proteases. Exploring the physiological role of each protease requires the identification of their natural substrates. However, the endogenous substrates of many of the human proteases are as yet unknown. Here we describe a new assay that addresses this problem. The assay, which easily can be automated, is based on the incubation of immobilized protein fractions, which may contain the natural substrate, with a defined protease. After concentrating the proteolytically released peptides by reversed-phase chromatography they are analyzed by tandem mass spectrometry and the substrates identified by database searching. The proof of principle in this study is demonstrated by incubating immobilized human plasma proteins with thrombin and by identifying by tandem mass spectrometry the fibrinopeptides, released by the action of thrombin from their natural substrate fibrinogen, in the reaction mixture.

KW - Blood Proteins

KW - Fibrinogen

KW - Humans

KW - Peptide Hydrolases

KW - Peptides

KW - Substrate Specificity

KW - Tandem Mass Spectrometry

KW - Thrombin

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1021/ac061482l

DO - 10.1021/ac061482l

M3 - SCORING: Journal article

C2 - 17263361

VL - 79

SP - 1251

EP - 1255

JO - ANAL CHEM

JF - ANAL CHEM

SN - 0003-2700

IS - 3

ER -