title: PITPs as targets for selectively interfering with phosphoinositide signaling in cells.
abstract: Sec14-like phosphatidylinositol transfer proteins (PITPs) integrate diverse territories of intracellular lipid metabolism with stimulated phosphatidylinositol-4-phosphate production and are discriminating portals for interrogating phosphoinositide signaling. Yet, neither Sec14-like PITPs nor PITPs in general have been exploited as targets for chemical inhibition for such purposes. Herein, we validate what is to our knowledge the first small-molecule inhibitors (SMIs) of the yeast PITP Sec14. These SMIs are nitrophenyl(4-(2-methoxyphenyl)piperazin-1-yl)methanones (NPPMs) and are effective inhibitors in vitro and in vivo. We further establish that Sec14 is the sole essential NPPM target in yeast and that NPPMs exhibit exquisite targeting specificities for Sec14 (relative to related Sec14-like PITPs), propose a mechanism for how NPPMs exert their inhibitory effects and demonstrate that NPPMs exhibit exquisite pathway selectivity in inhibiting phosphoinositide signaling in cells. These data deliver proof of concept that PITP-directed SMIs offer new and generally applicable avenues for intervening with phosphoinositide signaling pathways with selectivities superior to those afforded by contemporary lipid kinase-directed strategies.
authors.affiliation: 1] Department of Molecular and Cellular Medicine, Texas A&M University College Station, Texas, USA. [2] Department of Biochemistry and Biophysics, Texas A&M University College Station, Texas, USA. [3] Department of Chemistry, Texas A&M University College Station, Texas, USA. [4] Department of Cell and Developmental Biology, Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Aaron H
authors.initials: AH
authors.lastname: Nile
authors.affiliation: 1] Department of Molecular and Cellular Medicine, Texas A&M University College Station, Texas, USA. [2] Department of Biochemistry and Biophysics, Texas A&M University College Station, Texas, USA. [3] Department of Chemistry, Texas A&M University College Station, Texas, USA. [4] Laboratory for Molecular Modeling, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Tripathi
authors.affiliation: 1] Department of Molecular and Cellular Medicine, Texas A&M University College Station, Texas, USA. [2] Department of Biochemistry and Biophysics, Texas A&M University College Station, Texas, USA. [3] Department of Chemistry, Texas A&M University College Station, Texas, USA.
authors.forename: Peihua
authors.initials: P
authors.lastname: Yuan
authors.affiliation: 1] Department of Molecular and Cellular Medicine, Texas A&M University College Station, Texas, USA. [2] Department of Biochemistry and Biophysics, Texas A&M University College Station, Texas, USA. [3] Department of Chemistry, Texas A&M University College Station, Texas, USA.
authors.forename: Carl J
authors.initials: CJ
authors.lastname: Mousley
authors.affiliation: Department of Biochemistry, Stanford Genome Technology Center, Stanford University, Palo Alto, California, USA.
authors.forename: Sundari
authors.initials: S
authors.lastname: Suresh
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Iain M
authors.initials: IM
authors.lastname: Wallace
authors.affiliation: Department of Cell and Developmental Biology, Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Sweety D
authors.initials: SD
authors.lastname: Shah
authors.affiliation: Laboratory for Molecular Modeling, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Denise Teotico
authors.initials: DT
authors.lastname: Pohlhaus
authors.affiliation: R.L. Juliano Structural Bioinformatics Core University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Temple
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Guri
authors.initials: G
authors.lastname: Giaever
authors.affiliation: Laboratory for Molecular Modeling, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Tropsha
authors.affiliation: Department of Biochemistry, Stanford Genome Technology Center, Stanford University, Palo Alto, California, USA.
authors.forename: Ronald W
authors.initials: RW
authors.lastname: Davis
authors.affiliation: Department of Biochemistry, Stanford Genome Technology Center, Stanford University, Palo Alto, California, USA.
authors.forename: Robert P
authors.initials: RP
authors.lastname: St Onge
authors.affiliation: 1] Department of Molecular and Cellular Medicine, Texas A&M University College Station, Texas, USA. [2] Department of Biochemistry and Biophysics, Texas A&M University College Station, Texas, USA. [3] Department of Chemistry, Texas A&M University College Station, Texas, USA. [4] Department of Cell and Developmental Biology, Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Vytas A
authors.initials: VA
authors.lastname: Bankaitis
mesh_terms: D010716:Phosphatidylinositols / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction; D013329:Structure-Activity Relationship
chemical_list: D010716:Phosphatidylinositols; D049294:Phospholipid Transfer Proteins