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Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum

Serine palmitoyltransferase (SPT) is a key enzyme of sphingolipid biosynthesis, which catalyzes the pyridoxal-5′-phosphate–dependent decarboxylative condensation reaction of l-serine (l-Ser) and palmitoyl-CoA (PalCoA) to form 3-ketodihydrosphingosine called long chain base (LCB). SPT is also able to...

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Autores principales: Ikushiro, Hiroko, Murakami, Taiki, Takahashi, Aya, Katayama, Asuka, Sawai, Taiki, Goto, Haruna, Koolath, Sajeer, Murai, Yuta, Monde, Kenji, Miyahara, Ikuko, Kamiya, Nobuo, Yano, Takato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10196870/
https://www.ncbi.nlm.nih.gov/pubmed/37030501
http://dx.doi.org/10.1016/j.jbc.2023.104684
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author Ikushiro, Hiroko
Murakami, Taiki
Takahashi, Aya
Katayama, Asuka
Sawai, Taiki
Goto, Haruna
Koolath, Sajeer
Murai, Yuta
Monde, Kenji
Miyahara, Ikuko
Kamiya, Nobuo
Yano, Takato
author_facet Ikushiro, Hiroko
Murakami, Taiki
Takahashi, Aya
Katayama, Asuka
Sawai, Taiki
Goto, Haruna
Koolath, Sajeer
Murai, Yuta
Monde, Kenji
Miyahara, Ikuko
Kamiya, Nobuo
Yano, Takato
author_sort Ikushiro, Hiroko
collection PubMed
description Serine palmitoyltransferase (SPT) is a key enzyme of sphingolipid biosynthesis, which catalyzes the pyridoxal-5′-phosphate–dependent decarboxylative condensation reaction of l-serine (l-Ser) and palmitoyl-CoA (PalCoA) to form 3-ketodihydrosphingosine called long chain base (LCB). SPT is also able to metabolize l-alanine (l-Ala) and glycine (Gly), albeit with much lower efficiency. Human SPT is a membrane-bound large protein complex containing SPTLC1/SPTLC2 heterodimer as the core subunits, and it is known that mutations of the SPTLC1/SPTLC2 genes increase the formation of deoxy-type of LCBs derived from l-Ala and Gly to cause some neurodegenerative diseases. In order to study the substrate recognition of SPT, we examined the reactivity of Sphingobacterium multivorum SPT on various amino acids in the presence of PalCoA. The S. multivorum SPT could convert not only l-Ala and Gly but also l-homoserine, in addition to l-Ser, into the corresponding LCBs. Furthermore, we obtained high-quality crystals of the ligand-free form and the binary complexes with a series of amino acids, including a nonproductive amino acid, l-threonine, and determined the structures at 1.40 to 1.55 Å resolutions. The S. multivorum SPT accommodated various amino acid substrates through subtle rearrangements of the active-site amino acid residues and water molecules. It was also suggested that non-active-site residues mutated in the human SPT genes might indirectly influence the substrate specificity by affecting the hydrogen-bonding networks involving the bound substrate, water molecules, and amino acid residues in the active site of this enzyme. Collectively, our results highlight SPT structural features affecting substrate specificity for this stage of sphingolipid biosynthesis.
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spelling pubmed-101968702023-05-20 Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum Ikushiro, Hiroko Murakami, Taiki Takahashi, Aya Katayama, Asuka Sawai, Taiki Goto, Haruna Koolath, Sajeer Murai, Yuta Monde, Kenji Miyahara, Ikuko Kamiya, Nobuo Yano, Takato J Biol Chem Research Article Serine palmitoyltransferase (SPT) is a key enzyme of sphingolipid biosynthesis, which catalyzes the pyridoxal-5′-phosphate–dependent decarboxylative condensation reaction of l-serine (l-Ser) and palmitoyl-CoA (PalCoA) to form 3-ketodihydrosphingosine called long chain base (LCB). SPT is also able to metabolize l-alanine (l-Ala) and glycine (Gly), albeit with much lower efficiency. Human SPT is a membrane-bound large protein complex containing SPTLC1/SPTLC2 heterodimer as the core subunits, and it is known that mutations of the SPTLC1/SPTLC2 genes increase the formation of deoxy-type of LCBs derived from l-Ala and Gly to cause some neurodegenerative diseases. In order to study the substrate recognition of SPT, we examined the reactivity of Sphingobacterium multivorum SPT on various amino acids in the presence of PalCoA. The S. multivorum SPT could convert not only l-Ala and Gly but also l-homoserine, in addition to l-Ser, into the corresponding LCBs. Furthermore, we obtained high-quality crystals of the ligand-free form and the binary complexes with a series of amino acids, including a nonproductive amino acid, l-threonine, and determined the structures at 1.40 to 1.55 Å resolutions. The S. multivorum SPT accommodated various amino acid substrates through subtle rearrangements of the active-site amino acid residues and water molecules. It was also suggested that non-active-site residues mutated in the human SPT genes might indirectly influence the substrate specificity by affecting the hydrogen-bonding networks involving the bound substrate, water molecules, and amino acid residues in the active site of this enzyme. Collectively, our results highlight SPT structural features affecting substrate specificity for this stage of sphingolipid biosynthesis. American Society for Biochemistry and Molecular Biology 2023-04-07 /pmc/articles/PMC10196870/ /pubmed/37030501 http://dx.doi.org/10.1016/j.jbc.2023.104684 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Ikushiro, Hiroko
Murakami, Taiki
Takahashi, Aya
Katayama, Asuka
Sawai, Taiki
Goto, Haruna
Koolath, Sajeer
Murai, Yuta
Monde, Kenji
Miyahara, Ikuko
Kamiya, Nobuo
Yano, Takato
Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title_full Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title_fullStr Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title_full_unstemmed Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title_short Structural insights into the substrate recognition of serine palmitoyltransferase from Sphingobacterium multivorum
title_sort structural insights into the substrate recognition of serine palmitoyltransferase from sphingobacterium multivorum
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10196870/
https://www.ncbi.nlm.nih.gov/pubmed/37030501
http://dx.doi.org/10.1016/j.jbc.2023.104684
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