title: Temporally controlled targeting of 4-hydroxynonenal to specific proteins in living cells.
abstract: In-depth chemical understanding of complex biological processes hinges upon the ability to systematically perturb individual systems. However, current approaches to study impacts of biologically relevant reactive small molecules involve bathing of the entire cell or isolated organelle with excess amounts, leading to off-target effects. The resultant lack of biochemical specificity has plagued our understanding of how biological electrophiles mediate signal transduction or regulate responses that confer defense mechanisms to cellular electrophilic stress. Here we introduce a target-specific electrophile delivery platform that will ultimately pave the way to interrogate effects of reactive electrophiles on specific target proteins in cells. The new methodology is demonstrated by photoinducible targeted delivery of 4-hydroxynonenal (HNE) to the proteins Keap1 and PTEN. Covalent conjugation of the HNE-precursor to HaloTag fused to the target proteins enables directed HNE delivery upon photoactivation. The strategy provides proof of concept of selective delivery of reactive electrophiles to individual electrophile-responsive proteins in mammalian cells. It opens a new avenue enabling more precise determination of the pathophysiological consequences of HNE-induced chemical modifications on specific target proteins in cells.
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Xinqiang
authors.initials: X
authors.lastname: Fang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Fu
authors.forename: Marcus J C
authors.initials: MJ
authors.lastname: Long
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Haegele
authors.forename: Eva J
authors.initials: EJ
authors.lastname: Ge
authors.forename: Saba
authors.initials: S
authors.lastname: Parvez
authors.forename: Yimon
authors.initials: Y
authors.lastname: Aye
mesh_terms: D000447:Aldehydes / Q000008:administration & dosage*; D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D016503:Drug Delivery Systems; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D000072019:Kelch-Like ECH-Associated Protein 1; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D000447:Aldehydes; D047908:Intracellular Signaling Peptides and Proteins; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051059:PTEN Phosphohydrolase; C027576:4-hydroxy-2-nonenal