Assay of Cystathionine-γ-synthase (CaStr2p) activity determination. - Open Research Data - Bridge of Knowledge

Search

Assay of Cystathionine-γ-synthase (CaStr2p) activity determination.

Description

The study aimed to obtain conditions optimal for the activity of Candida albicans cystathionine-γ-synthase (CaStr2p) determination. The selection of appropriate reaction conditions included the identification of the reaction buffer and its pH, substrate, and enzyme concentrations. The activity of Str2p was measured by the detection of decrease of the substrate L-cysteine (L-Cys) via LC/MS system with pre-column derivatization. Mass spectra were recorded using an Agilent 6470A triple quadrupole (6470 Triple Quad LC/MS, Agilent Technologies, Waldbronn, Germany) LC/MS system (1260 Infinity II, Agilent Technologies,Waldbronn, Germany) with electrospray ionization source (ESI). L-Cys reacts with 5,5’-dithiobis(2-nitrobenzoic acid) (DTNB) to produce a L-Cys-TNB complex, that can be also detected by the diode array detector (HPLC-DAD). Optimal conditions for L-Cys-TNB complex detection included identification of mobile phase, elution profile, injection volume, and most suitable wavelengths for the detection have been identified.

This dataset contains the description of the methodology, and the set of selected chromatographs obtained for control as well as enzymatic reaction samples.

The studies were financially supported by the project OPUS 20 financed by the National Science Centre (No. UMO-2020/39/B/NZ7/01519).

Dataset file

Assay of CaStr2p activity determination.zip
641.2 kB, S3 ETag 1e417453e0da41a10b31a3bd44903696-1, downloads: 44
The file hash is calculated from the formula
hexmd5(md5(part1)+md5(part2)+...)-{parts_count} where a single part of the file is 512 MB in size.

Example script for calculation:
https://github.com/antespi/s3md5
download file Assay of CaStr2p activity determination.zip

File details

License:
Creative Commons: by 4.0 open in new tab
CC BY
Attribution
Raw data:
Data contained in dataset was not processed.

Details

Year of publication:
2024
Verification date:
2024-06-14
Dataset language:
English
Fields of science:
  • chemical sciences (Natural sciences)
DOI:
DOI ID 10.34808/st1j-4j69 open in new tab
Funding:
Verified by:
Gdańsk University of Technology

Keywords

References

Cite as

seen 83 times