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A spinach O-acetylserine(thiol)lyase homologue, SoCSaseLP, suppresses cysteine biosynthesis catalysed by other enzyme isoforms.
Biochim Open 2016; 2:24-32BO

Abstract

An enzyme, O-acetylserine(thiol)lyase (OASTL), also known as O-acetylserine sulfhydrylase or cysteine synthase (CSase), catalyses the incorporation of sulfide into O-acetylserine and produces cysteine. We previously identified a cDNA encoding an OASTL-like protein from Spinacia oleracea, (SoCSaseLP), but a recombinant SoCSaseLP produced in Escherichia coli did not show OASTL activity. The exon-intron structure of the SoCSaseLP gene shared conserved structures with other spinach OASTL genes. The SoCSaseLP and a Beta vulgaris homologue protein, KMT13462, comprise a unique clade in the phylogenetic tree of the OASTL family. Interestingly, when the SoCSaseLP gene was expressed in tobacco plants, total OASTL activity in tobacco leaves was reduced. This reduction in total OASTL activity was most likely caused by interference by SoCSaseLP with cytosolic OASTL. To investigate the possible interaction of SoCSaseLP with a spinach cytosolic OASTL isoform SoCSaseA, a pull-down assay was carried out. The recombinant glutathione S-transferase (GST)-SoCSaseLP fusion protein was expressed in E. coli together with the histidine-tagged SoCSaseA protein, and the protein extract was subjected to glutathione affinity chromatography. The histidine-tagged SoCSaseA was co-purified with the GST-SoCSaseLP fusion protein, indicating the binding of SoCSaseLP to SoCSaseA. Consistent with this interaction, the OASTL activity of the co-purified SoCSaseA was reduced compared with the activity of SoCSaseA that was purified on its own. These results strongly suggest that SoCSaseLP negatively regulates the activity of other cytosolic OASTL family members by direct interaction.

Authors+Show Affiliations

Graduate School of Horticulture, Chiba University, 648 Matsudo, Chiba 271-8510, Japan.Graduate School of Science and Technology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.Graduate School of Science and Technology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.Graduate School of Science and Technology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.Graduate School of Advanced Integration Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.Graduate School of Advanced Integration Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29632835

Citation

Noda, Miki, et al. "A Spinach O-acetylserine(thiol)lyase Homologue, SoCSaseLP, Suppresses Cysteine Biosynthesis Catalysed By Other Enzyme Isoforms." Biochimie Open, vol. 2, 2016, pp. 24-32.
Noda M, Nakamura M, Takamiya R, et al. A spinach O-acetylserine(thiol)lyase homologue, SoCSaseLP, suppresses cysteine biosynthesis catalysed by other enzyme isoforms. Biochim Open. 2016;2:24-32.
Noda, M., Nakamura, M., Takamiya, R., Tamura, T., Ito, T., & Kodama, H. (2016). A spinach O-acetylserine(thiol)lyase homologue, SoCSaseLP, suppresses cysteine biosynthesis catalysed by other enzyme isoforms. Biochimie Open, 2, pp. 24-32. doi:10.1016/j.biopen.2016.01.002.
Noda M, et al. A Spinach O-acetylserine(thiol)lyase Homologue, SoCSaseLP, Suppresses Cysteine Biosynthesis Catalysed By Other Enzyme Isoforms. Biochim Open. 2016;2:24-32. PubMed PMID: 29632835.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A spinach O-acetylserine(thiol)lyase homologue, SoCSaseLP, suppresses cysteine biosynthesis catalysed by other enzyme isoforms. AU - Noda,Miki, AU - Nakamura,Mika, AU - Takamiya,Ryuichi, AU - Tamura,Takashi, AU - Ito,Toshiyuki, AU - Kodama,Hiroaki, Y1 - 2016/02/08/ PY - 2015/12/11/received PY - 2016/01/31/accepted PY - 2018/4/11/entrez PY - 2016/2/8/pubmed PY - 2016/2/8/medline KW - CSC, cysteine synthase complex KW - CSase, cysteine synthase KW - Cysteine synthesis KW - GST, glutathione S-transferase KW - Negative regulation KW - O-acetylserine(thiol)lyase KW - OAS, O-acetylserine KW - OASTL, O-acetylserine(thiol)lyase KW - SAT, serine acetyltransferase KW - Spinach SP - 24 EP - 32 JF - Biochimie open JO - Biochim Open VL - 2 N2 - An enzyme, O-acetylserine(thiol)lyase (OASTL), also known as O-acetylserine sulfhydrylase or cysteine synthase (CSase), catalyses the incorporation of sulfide into O-acetylserine and produces cysteine. We previously identified a cDNA encoding an OASTL-like protein from Spinacia oleracea, (SoCSaseLP), but a recombinant SoCSaseLP produced in Escherichia coli did not show OASTL activity. The exon-intron structure of the SoCSaseLP gene shared conserved structures with other spinach OASTL genes. The SoCSaseLP and a Beta vulgaris homologue protein, KMT13462, comprise a unique clade in the phylogenetic tree of the OASTL family. Interestingly, when the SoCSaseLP gene was expressed in tobacco plants, total OASTL activity in tobacco leaves was reduced. This reduction in total OASTL activity was most likely caused by interference by SoCSaseLP with cytosolic OASTL. To investigate the possible interaction of SoCSaseLP with a spinach cytosolic OASTL isoform SoCSaseA, a pull-down assay was carried out. The recombinant glutathione S-transferase (GST)-SoCSaseLP fusion protein was expressed in E. coli together with the histidine-tagged SoCSaseA protein, and the protein extract was subjected to glutathione affinity chromatography. The histidine-tagged SoCSaseA was co-purified with the GST-SoCSaseLP fusion protein, indicating the binding of SoCSaseLP to SoCSaseA. Consistent with this interaction, the OASTL activity of the co-purified SoCSaseA was reduced compared with the activity of SoCSaseA that was purified on its own. These results strongly suggest that SoCSaseLP negatively regulates the activity of other cytosolic OASTL family members by direct interaction. SN - 2214-0085 UR - https://www.unboundmedicine.com/medline/citation/29632835/A_spinach_O_acetylserine_thiol_lyase_homologue_SoCSaseLP_suppresses_cysteine_biosynthesis_catalysed_by_other_enzyme_isoforms_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S2214-0085(16)00003-1 DB - PRIME DP - Unbound Medicine ER -