title: Covalent Modification of CDK2 by 4-Hydroxynonenal as a Mechanism of Inhibition of Cell Cycle Progression.
abstract: Oxidative stress is a contributing factor in a number of chronic diseases, including cancer, atherosclerosis, and neurodegenerative diseases. Lipid peroxidation that occurs during periods of oxidative stress results in the formation of lipid electrophiles, which can modify a multitude of proteins in the cell. 4-Hydroxy-2-nonenal (HNE) is one of the most well-studied lipid electrophiles and has previously been shown to arrest cells at the G1/S transition. Recently, proteomic data have shown that HNE is capable of covalently modifying CDK2, the kinase responsible for the G1/S transition. Here, we identify the sites adducted by HNE using recombinant CDK2 and show that HNE treatment suppresses the kinase activity of the enzyme. We further identify sites of adduction in HNE-treated intact human colorectal carcinoma cells (RKO) and show that HNE-dependent modification in cells is long-lived, disrupts CDK2 function, and correlates with a delay of progression of the cells into S-phase. We propose that adduction of CDK2 by HNE directly alters its activity, contributing to the cell cycle delay.
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, §Chemistry, and #Pharmacology, ‡Mass Spectrometry Research Center, ⊥Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Jeannie M
authors.initials: JM
authors.lastname: Camarillo
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, §Chemistry, and #Pharmacology, ‡Mass Spectrometry Research Center, ⊥Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Kristie L
authors.initials: KL
authors.lastname: Rose
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, §Chemistry, and #Pharmacology, ‡Mass Spectrometry Research Center, ⊥Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: James J
authors.initials: JJ
authors.lastname: Galligan
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, §Chemistry, and #Pharmacology, ‡Mass Spectrometry Research Center, ⊥Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Shu
authors.initials: S
authors.lastname: Xu
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, §Chemistry, and #Pharmacology, ‡Mass Spectrometry Research Center, ⊥Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D000447:Aldehydes / Q000737:chemistry* / Q000494:pharmacology*; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D051357:Cyclin-Dependent Kinase 2 / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008958:Models, Molecular; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: D000447:Aldehydes; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; C027576:4-hydroxy-2-nonenal