Glycosylinositol phosphoceramide-specific phospholipase D activity catalyzes transphosphatidylation

2019 ◽  
Vol 166 (5) ◽  
pp. 441-448 ◽  
Author(s):  
Rumana Yesmin Hasi ◽  
Makoto Miyagi ◽  
Katsuya Morito ◽  
Toshiki Ishikawa ◽  
Maki Kawai-Yamada ◽  
...  
Keyword(s):  
Chain Length ◽  
Phospholipase D ◽  
Secondary Alcohol ◽  
Primary Alcohol ◽  
Tertiary Alcohol ◽  
Head Group ◽  
Polar Head Group ◽  
Polar Head ◽  
A Chain ◽  
Sugar Chains

Abstract Glycosylinositol phosphoceramide (GIPC) is the most abundant sphingolipid in plants and fungi. Recently, we detected GIPC-specific phospholipase D (GIPC-PLD) activity in plants. Here, we found that GIPC-PLD activity in young cabbage leaves catalyzes transphosphatidylation. The available alcohol for this reaction is a primary alcohol with a chain length below C4. Neither secondary alcohol, tertiary alcohol, choline, serine nor glycerol serves as an acceptor for transphosphatidylation of GIPC-PLD. We also found that cabbage GIPC-PLD prefers GIPC containing two sugars. Neither inositol phosphoceramide, mannosylinositol phosphoceramide nor GIPC with three sugar chains served as substrate. GIPC-PLD will become a useful catalyst for modification of polar head group of sphingophospholipid.

Mixed Micelles Containing Sodium Laurate: Effect of Chain Length, Polar Head Group, and Added Salt

2012 ◽  
Vol 49 (6) ◽  
pp. 488-493 ◽  
Author(s):  
Rie Kakehashi ◽  
Motohiro Shizuma ◽  
Shingo Yamamura
Keyword(s):  
Chain Length ◽  
Mixed Micelles ◽  
Sodium Laurate ◽  
Head Group ◽  
Polar Head Group ◽  
Polar Head ◽  
Added Salt

Mixed micelles containing sodium oleate: the effect of the chain length and the polar head group

2004 ◽  
Vol 279 (1) ◽  
pp. 253-258 ◽  
Author(s):  
Rie Kakehashi ◽  
Motohiro Shizuma ◽  
Shingo Yamamura ◽  
Tokuji Takeda
Keyword(s):  
Chain Length ◽  
Sodium Oleate ◽  
Mixed Micelles ◽  
Head Group ◽  
Polar Head Group ◽  
Polar Head

scholarly journals Structures of an engineered phospholipase D with specificity for secondary alcohol transphosphatidylation: Insights into plasticity of substrate binding and activation

2021 ◽  
Author(s):  
Ariela Samantha ◽  
Jasmina Damnjanović ◽  
Yugo Iwasaki ◽  
Hideo Nakano ◽  
Alice Vrielink
Keyword(s):  
Phospholipase D ◽  
Catalytic Mechanism ◽  
Inositol Phosphate ◽  
Substrate Binding ◽  
Secondary Alcohol ◽  
Molecular Size ◽  
Underlying Mechanism ◽  
Head Group ◽  
Wild Type ◽  
Primary Alcohols

Phospholipase D (PLD) is an enzyme useful for the enzymatic modification of phospholipids.  In the presence of primary alcohols, the enzyme catalyses transphosphatidylation of the head group of phospholipid substrates to synthesize a modified phospholipid product.  However, the enzyme is specific for primary alcohols and thus the limitation of the molecular size of the acceptor compounds has restricted the type of phospholipid species that can be synthesised.  An engineered variant of PLD from Streptomycesantibioticus termed TNYR SaPLD was developed capable of synthesizing 1-phosphatidylinositol with positional specificity of up to 98%. To gain a better understanding of the substrate binding features of the TNYR SaPLD, crystal structures have been determined for the free enzyme and its complexes with phosphate, phosphatidic acid and 1-inositol phosphate.  Comparisons of these structures with the wild-type SaPLD show a larger binding site able to accommodate a bulkier secondary alcohol substrate as well as changes to the position of a flexible surface loop proposed to be involved in substrate recognition.  The complex of the active TNYR SaPLD with 1-inositol phosphate reveals a covalent intermediate adduct with the ligand bound to H442 rather than to H168, the proposed nucleophile in the wild type enzyme.  This structural feature suggests that the enzyme exhibits plasticity of the catalytic mechanism different from what has been reported to date for PLDs.  These structural studies provide insights into the underlying mechanism that governs the recognition of myo-inositol by TNYR SaPLD, and an important foundation for further studies of the catalytic mechanism.

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