scholarly journals A Liquid Chromatography/Mass Spectrometry Method for Screening Disulfide Tethering Fragments

2017 ◽  
Vol 23 (2) ◽  
pp. 183-192 ◽  
Author(s):  
Kenneth K. Hallenbeck ◽  
Julia L. Davies ◽  
Connie Merron ◽  
Pierce Ogden ◽  
Eline Sijbesma ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Data Collection ◽  
Screening Method ◽  
Liquid Chromatography Mass Spectrometry ◽  
Compound Libraries ◽  
Chromatography Mass Spectrometry ◽  
Molecular Masses ◽  
Signal Significance ◽  
Covalent Adducts

We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)–based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a percent bound value for each compound and identifies false positives by calculating significance of detected masses (signal significance). An example pipeline is available through Biovia’s ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1 to 2 days.

scholarly journals A Liquid Chromatography-Mass Spectrometry Method for Screening Disulfide Tethering Fragments

2017 ◽  
Author(s):  
Kenneth K Hallenbeck ◽  
Julia L. Davies ◽  
Connie Merron ◽  
Pierce Ogden ◽  
Eline Sijbesma ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Data Collection ◽  
Screening Method ◽  
Liquid Chromatography Mass Spectrometry ◽  
Compound Libraries ◽  
Chromatography Mass Spectrometry ◽  
Molecular Masses ◽  
Signal Significance ◽  
Covalent Adducts

AbstractWe report the refinement of a high-throughput, liquid-chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 90 seconds per sample balances speed and reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a % bound value for each compound and detects false-positives by calculating significance of detected masses (‘signal significance’). An example pipeline has been made available through Biovia’s ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1–2 days.

Detection of Covalent Adducts to Cytochrome P450 3A4 Using Liquid Chromatography Mass Spectrometry

2004 ◽  
Vol 17 (10) ◽  
pp. 1356-1361 ◽  
Author(s):  
Kevin P. Bateman ◽  
Jennifer Baker ◽  
Mark Wilke ◽  
Justin Lee ◽  
Tammy LeRiche ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cytochrome P450 ◽  
Liquid Chromatography ◽  
Cytochrome P450 3A4 ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Covalent Adducts

scholarly journals Screening for Sulfur Compounds by Molybdenum-Catalyzed Oxidation Combined with Liquid Chromatography-Mass Spectrometry

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 240 ◽  
Author(s):  
Hirotaka Matsuo ◽  
Yu Hanamure ◽  
Rei Miyano ◽  
Yōko Takahashi ◽  
Satoshi Ōmura ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Secondary Alcohol ◽  
Screening Method ◽  
Sulfur Compounds ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Actinomycete Strain ◽  
Neutral Conditions ◽  
Catalyzed Oxidation

The molybdenum (Mo)-catalyzed oxidation of sulfide under neutral conditions yields sulfone. This reaction proceeds more smoothly than olefin epoxidation and primary or secondary alcohol oxidation. In this study, Mo-catalyzed oxidation was used to screen for sulfur compounds (named “MoS-screening”) in microbial broths by liquid chromatography-mass spectrometry (LC/MS). To demonstrate proof-of-concept, known sulfur microbial compounds were successfully identified from a mixture of non-sulfur microbial compounds as sulfinyl or sulfonyl products of Mo-catalyzed oxidation. Then our MoS-screening method was used to screen 300 samples of microbial broth for sulfur compounds. One of the identified compounds was a kitasetaline-containing N-acetyl cysteine moiety produced by an actinomycete strain. These results demonstrate the potential of MoS-screening in the search for new sulfur compounds from microbial sources.

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