Structure-Functional Examination of Cysteine Synthase A (CysK) from Limosilactobacillus reuteri LR1
Full article
| Journal |
International Journal of Molecular Sciences
ISSN: 1661-6596
, E-ISSN: 1422-0067
|
| Output data |
Year: 2026,
Volume: 27,
Number: 1,
Article number
: 327,
Pages count
:
DOI:
10.3390/ijms27010327
|
| Authors |
Pometun Anastasia A.
1,2
,
Les Evgenii K.
1,3
,
Chernobrovkina Alla V.
3
,
Gorbovskaia Anastasiia V.
1,3
,
Chikurova Natalia Yu
1,3
,
Loginova Anastasia A.
1,3
,
Antipov Alexey N.
1
,
Mordkovich Nadezhda N.
1
,
Shaposhnikov Leonid A.
1
,
Savin Svyatoslav S.
1,3
,
Kleymenov Sergey Yu
1,4
,
Matyuta Ilya O.
1
,
Boyko Konstantin M.
1
,
Minyaev Mikhail E.
5
,
Hushpulian Dmitry M.
1
,
Pometun Evgenii V.
6
,
Tishkov Vladimir I.
1,3
|
| Affiliations |
| 1 |
Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 33, bld. 2 Leninsky Avenue, 119071 Moscow, Russia
|
| 2 |
Institute of Medicine, Peoples’ Friendship, University of Russia Named After Patrice Lumumba, Miklouho-Maklaya, 8, 117198 Moscow, Russia
|
| 3 |
Department of Chemistry, Lomonosov Moscow State University, Leninskie Gory, 1-3, 119991 Moscow, Russia
|
| 4 |
Koltzov Institute of Developmental Biology of Russian Academy of Sciences, Vavilova, 26, 119334 Moscow, Russia
|
| 5 |
N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Avenue, 47, 119991 Moscow, Russia
|
| 6 |
Federal State Autonomous Educational Institution of Higher Education, I.M. Sechenov First Moscow State Medical University (Sechenovskiy University), Trubetskaya St., 8, Building 2, 119048 Moscow, Russia
|
|
Funding (1)
This study presents a comprehensive analysis of cysteine synthase A (CysK) from Limosilactobacillus reuteri LR1 (LreCysK), an enzyme involved in the biosynthesis of L-cysteine. This protein supports crucial cellular functions such as sulfur metabolism, antioxidant defense, detoxification, and protein synthesis. Previously, the gene encoding LreCysK was cloned, and the enzyme with His-tag on the N-terminus was obtained in active and soluble form. Here, kinetic parameters of the enzyme were determined by the previously developed high-pressure liquid chromatography (HPLC) and ninhydrin methods. It was found that LreCysK has similar KMOAS and kcat as CysKs from Escherichia coli and from the model plant Arabidopsis thaliana. The thermal stability of LreCysK was studied using differential scanning calorimetry. It was revealed that the melting point of the enzyme increases to almost 90°C when Pyridoxal-5 phosphate (PLP) is added, indicating that the stability of the enzyme complex with PLP is relatively high. Structural studies revealed that LreCysK is a dimer, and its active site is similar to those of other enzymes, but exhibits some features characteristic of lactobacilli CysKs (GISA), as well as unique residues, such as Ile50. Also, the potential biotechnological applications of LreCysK are discussed. These findings enhance our understanding of LreCysK’s biochemical versatility and its potential applications in biotechnology and medicine.