Phosphatidylserine synthesis in Saccharomyces cerevisiae. Purification and characterization of membrane-associated phosphatidylserine synthase.

Bae-Lee, Myongsuk; Carman, George M.

doi:10.1016/s0021-9258(18)90592-2

Cited by 124 publications

(33 citation statements)

References 34 publications

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“…This reaction is catalyzed by CDP-DAG alcohol O-phosphatidyl transferases (CDP-AP, EC 2.7.8.8) and leads to the synthesis of a lipid molecule and the release of CMP. In some Gram-negative bacteria, the enzymes are peripheral membrane proteins and are associated with the soluble ribosomal fraction 13 – 15 , whereas in Gram-positive bacteria, lower eukaryotes, and plants the enzymes are integral membrane proteins 16 , 17 . The hydrophobic chain of the substrates, which are geranyl groups (CDP-archaeols) in archaea and acyl groups (CDP-DAG) in bacteria or higher eukaryotes, has no obvious effect on CDP-AP activity 10 , 18 , 19 .…”

Section: Introductionmentioning

confidence: 99%

Crystal structures of phosphatidyl serine synthase PSS reveal the catalytic mechanism of CDP-DAG alcohol O-phosphatidyl transferases

et al. 2021

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Phospholipids are the major components of the membrane in all type of cells and organelles. They also are critical for cell metabolism, signal transduction, the immune system and other critical cell functions. The biosynthesis of phospholipids is a complex multi-step process with high-energy intermediates. Several enzymes in different metabolic pathways are involved in the initial phospholipid synthesis and its subsequent conversion. While the “Kennedy pathway” is the main pathway in mammalian cells, in bacteria and lower eukaryotes the precursor CDP-DAG is used in the de novo pathway by CDP-DAG alcohol O-phosphatidyl transferases to synthetize the basic lipids. Here we present the high-resolution structures of phosphatidyl serine synthase from Methanocaldococcus jannaschii crystallized in four different states. Detailed structural and functional analysis of the different structures allowed us to identify the substrate binding site and show how CDP-DAG, serine and two essential metal ions are bound and oriented relative to each other. In close proximity to the substrate binding site, two anions were identified that appear to be highly important for the reaction. The structural findings were confirmed by functional activity assays and suggest a model for the catalytic mechanism of CDP-DAG alcohol O-phosphatidyl transferases, which synthetize the phospholipids essential for the cells.

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Section: Introductionmentioning

confidence: 99%

Crystal structures of phosphatidyl serine synthase PSS reveal the catalytic mechanism of CDP-DAG alcohol O-phosphatidyl transferases

et al. 2021

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“…Somatic cells synthesize PS by headgroup exchange between PE or PC with L-serine (Stone and Vance, 2000). Bacteria (Kanfer and Kennedy, 1962) and most fungi (Bae-Lee and Carman, 1984) utilize the prokaryotic pathway where L-serine displaces CMP from CDP-DAG to form PS. Psd, which is highly homologous between eukaryotes and prokaryotes (Voelker, 1997), catalyzes the decarboxylation of PS to PE (Kanfer and Kennedy, 1964).…”

Section: Current State Of the Bacterial Kennedy Pathwaymentioning

confidence: 99%

“…PssA's in Gram-negative bacteria including E. coli Kennedy, 1972, 1974) are unique in that they are not associated with the membrane in cell free extracts but are tightly associated with ribosomes (Dutt and Dowhan, 1977). In Bacillus subtilis (Okada et al, 1994), Bacillus licheniformis (Dutt and Dowhan, 1981) and S. cerevisiae (Bae-Lee and Carman, 1984), the enzyme is an integral membrane protein requiring a divalent cation for activity, which the E. coli enzyme does not. The B. subtilis enzyme can fully substitute for the E. coli enzyme when expressed in a pssA null strain (Okada et al, 1994).…”

Section: Ps Synthasementioning

confidence: 99%

Eugene P. Kennedy’s Legacy: Defining Bacterial Phospholipid Pathways and Function

Dowhan

Bogdanov

2021

Front. Mol. Biosci.

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In the 1950’s and 1960’s Eugene P. Kennedy laid out the blueprint for phospholipid biosynthesis in somatic cells and Escherichia coli, which have been coined the Kennedy Pathways for phospholipid biosynthesis. His research group continued to make seminal contributions in the area of phospholipids until his retirement in the early 1990’s. During these years he mentored many young scientists that continued to build on his early discoveries and who also mentored additional scientists that continue to make important contributions in areas related to phospholipids and membrane biogenesis. This review will focus on the initial E. coli Kennedy Pathways and how his early contributions have laid the foundation for our current understanding of bacterial phospholipid genetics, biochemistry and function as carried on by his scientific progeny and others who have been inspired to study microbial phospholipids.

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“…It was funded with a first-year direct cost budget of $37,295. Within the next few years, we purified the yeast PI synthase ( 42 ), PS synthase ( 43 , 44 ), and CDP–DAG synthase ( 45 ) using a modified version of the CDP–DAG–Sepharose affinity resin. This same grant, which received a merit award in 2010, was funded from 1980 to 2020.…”

Section: The Research That Shaped My Careermentioning

confidence: 99%

“…4 ). His work on the purification of PI synthase ( 42 ) and its reconstitution into liposomes ( 107 ) laid the groundwork for our studies on the PS synthase ( 44 , 108 ) and CDP–DAG synthase ( 45 ). Mike Homann’s contributions led to the initial purification and characterization of PA phosphatase by Yi-Ping Lin.…”

Section: The Mentors Mentees and Collaborators That Helped Shape My Careermentioning

confidence: 99%

Lipid metabolism has been good to me

Carman

2021

Journal of Biological Chemistry

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This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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Phosphatidylserine synthesis in Saccharomyces cerevisiae. Purification and characterization of membrane-associated phosphatidylserine synthase.

Cited by 124 publications

References 34 publications

Crystal structures of phosphatidyl serine synthase PSS reveal the catalytic mechanism of CDP-DAG alcohol O-phosphatidyl transferases

Crystal structures of phosphatidyl serine synthase PSS reveal the catalytic mechanism of CDP-DAG alcohol O-phosphatidyl transferases

Eugene P. Kennedy’s Legacy: Defining Bacterial Phospholipid Pathways and Function

Lipid metabolism has been good to me

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