Microbe Engineering of Guaia-6,10(14)-diene as a Building Block for the Semisynthetic Production of Plant-Derived (−)-Englerin A

Herein, we report the semisynthetic production of the potent transient receptor potential canonical (TRPC) channel agonist (−)-englerin A (EA), using guaia-6,10(14)-diene as the starting material. Guaia-6,10(14)-diene was systematically engineered in Escherichia coli and Saccharomyces cerevisiae using the CRISPR/Cas9 system and produced with high titers. This provided us the opportunity to execute a concise chemical synthesis of EA and the two related guaianes (−)-oxyphyllol and (+)-orientalol E. The potentially scalable approach combines the advantages of synthetic biology and chemical synthesis and provides an efficient and economical method for producing EA as well as its analogs.

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TRPC5 Does Not Cause or Aggravate Glomerular Disease

Journal of the American Society of Nephrology ◽

10.1681/asn.2017060682 ◽

2017 ◽

Vol 29 (2) ◽

pp. 409-415 ◽

Cited By ~ 20

Author(s):

Xuexiang Wang ◽

Ranadheer R. Dande ◽

Hao Yu ◽

Beata Samelko ◽

Rachel E. Miller ◽

...

Keyword(s):

Transient Receptor Potential ◽

Calcium Influx ◽

Receptor Potential ◽

Transient Receptor Potential Channel ◽

Transgenic Mouse Models ◽

Cytoskeletal Remodeling ◽

Englerin A ◽

Transient Receptor ◽

Ion Pore

Transient receptor potential channel 5 (TRPC5) is highly expressed in brain and kidney and mediates calcium influx and promotes cell migration. In the kidney, loss of TRPC5 function has been reported to benefit kidney filter dynamics by balancing podocyte cytoskeletal remodeling. However, in vivo gain-in-function studies of TRPC5 with respect to kidney function have not been reported. To address this gap, we developed two transgenic mouse models on the C57BL/6 background by overexpressing either wild-type TRPC5 or a TRPC5 ion-pore mutant. Compared with nontransgenic controls, neither transgenic model exhibited an increase in proteinuria at 8 months of age or a difference in LPS-induced albuminuria. Moreover, activation of TRPC5 by Englerin A did not stimulate proteinuria, and inhibition of TRPC5 by ML204 did not significantly lower the level of LPS-induced proteinuria in any group. Collectively, these data suggest that the overexpression or activation of the TRPC5 ion channel does not cause kidney barrier injury or aggravate such injury under pathologic conditions.

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Structure of the ancestral TRPY1 channel from Saccharomyces cerevisiae reveals mechanisms of modulation by lipids and calcium

10.1101/2020.10.12.336495 ◽

2020 ◽

Author(s):

Tofayel Ahmed ◽

Collin R. Nisler ◽

Edwin C. Fluck ◽

Marcos Sotomayor ◽

Vera Y. Moiseenkova-Bell

Keyword(s):

Saccharomyces Cerevisiae ◽

Transient Receptor Potential ◽

Transmembrane Domain ◽

Trp Channels ◽

Receptor Potential ◽

Lipid Binding ◽

Channel Modulation ◽

Channel Inhibition ◽

Transient Receptor ◽

Dynamics Simulations

ABSTRACTTransient Receptor Potential (TRP) channels have evolved in eukaryotes to control various cellular functions in response to a wide variety of chemical and physical stimuli. This large and diverse family of channels emerged in fungi as mechanosensitive osmoregulators. The Saccharomyces cerevisiae vacuolar TRP yeast 1 (TRPY1) is the most studied TRP channel from fungi, but the molecular details of channel modulation remain elusive so far. Here, we describe the full-length cryo-electron microscopy structure of TRPY1 at 3.1 Å resolution. The structure reveals a distinctive architecture for TRPY1 among all eukaryotic TRP channels with an evolutionarily conserved and archetypical transmembrane domain, but distinct structural folds for the cytosolic N- and C-termini. We identified the inhibitory phosphatidylinositol 3-phosphate (PI(3)P) lipid binding site, which sheds light into the lipid modulation of TRPY1 in the vacuolar membrane. The structure also exhibited two Ca2+-binding sites: one in the cytosolic side, implicated in channel activation, and the other in the vacuolar lumen side, involved in channel inhibition. These findings, together with data from molecular dynamics simulations, provide structural insights into the basis of TRPY1 channel modulation by lipids and Ca2+.

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Englerin A-sensing charged residues for transient receptor potential canonical 5 channel activation

Korean Journal of Physiology and Pharmacology ◽

10.4196/kjpp.2019.23.3.191 ◽

2019 ◽

Vol 23 (3) ◽

pp. 191 ◽

Cited By ~ 1

Author(s):

SeungJoo Jeong ◽

Juyeon Ko ◽

Minji Kim ◽

Ki Chul Park ◽

Eunice Yon June Park ◽

...

Keyword(s):

Transient Receptor Potential ◽

Receptor Potential ◽

Channel Activation ◽

Transient Receptor Potential Canonical ◽

Englerin A ◽

Charged Residues ◽

Transient Receptor

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Inositol lipids and TRPC channel activation

Biochemical Society Symposium ◽

10.1042/bss2007c04 ◽

2007 ◽

Vol 74 ◽

pp. 37-45 ◽

Cited By ~ 12

Author(s):

James W. Putney

Keyword(s):

Plasma Membrane ◽

Phospholipase C ◽

Transient Receptor Potential ◽

Calcium Signalling ◽

Receptor Potential ◽

Breakdown Product ◽

Channel Activation ◽

Inositol Lipids ◽

Transient Receptor ◽

Secondary Mechanism

The original hypothesis put forth by Bob Michell in his seminal 1975 review held that inositol lipid breakdown was involved in the activation of plasma membrane calcium channels or ‘gates’. Subsequently, it was demonstrated that while the interposition of inositol lipid breakdown upstream of calcium signalling was correct, it was predominantly the release of Ca2+ that was activated, through the formation of Ins(1,4,5)P3. Ca2+ entry across the plasma membrane involved a secondary mechanism signalled in an unknown manner by depletion of intracellular Ca2+ stores. In recent years, however, additional non-store-operated mechanisms for Ca2+ entry have emerged. In many instances, these pathways involve homologues of the Drosophila trp (transient receptor potential) gene. In mammalian systems there are seven members of the TRP superfamily, designated TRPC1–TRPC7, which appear to be reasonably close structural and functional homologues of Drosophila TRP. Although these channels can sometimes function as store-operated channels, in the majority of instances they function as channels more directly linked to phospholipase C activity. Three members of this family, TRPC3, 6 and 7, are activated by the phosphoinositide breakdown product, diacylglycerol. Two others, TRPC4 and 5, are also activated as a consequence of phospholipase C activity, although the precise substrate or product molecules involved are still unclear. Thus the TRPCs represent a family of ion channels that are directly activated by inositol lipid breakdown, confirming Bob Michell's original prediction 30 years ago.

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