https://www.galliganlab.com/welcome daily 0.75 2014-04-23 https://www.galliganlab.com/projects daily 0.75 2018-07-24 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1530912640085-E71KYRLVWGTB7EE8CVVO/Picture1.jpg Projects Histones are modified by MGO. (A) Western blotting demonstrates complete knockout of GLO1. (B) GLO1-/- cells lack any measurable GLO1 activity. (C) Western blotting of chromatin fractions using isoform-specific MG-H antibodies (courtesy of Dr. David Spiegel) reveals histones as targets for modification with markedly increased levels observed in GLO1-/- cells treated with electrophile. Due to a lack of measurable MG-H2 protein adduction, chromatin was treated with 5 mM MGO for 6 h (‘Control’) to serve as a positive control for MGO modification. (D,E) RNA-Seq reveals transcripts altered by MGO. Venn diagrams displaying significantly decreased or increased protein coding transcripts in GLO1-/- cells treated with either vehicle, 50 µM MGO, or 500 µM MGO compared with WT vehicle control. WT cells were also analyzed, revealing minimal alterations in gene expression, likely a result of the rapid metabolism of MGO by GLO1. Collectively, these data demonstrate that MGO mediates alterations in gene expression, which may be attributed, in part, to the covalent modification of histones. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1531244823990-M1LN6LN1WG5WKZAARTXF/Test.jpg Projects Methylglyoxal (MGO) is generated as a by-product of glycolysis (~0.1% - 0.4% of the total glycolytic flux). Under physiologic conditions, MGO is detoxified by the glyoxalase system, which is comprised of two enzymes (glyoxalase 1, GLO1, and GLO2). When this system is compromised (e.g. diabetes), concentrations of MGO are elevated and it can adduct proteins and DNA, yielding stable, long-lived adducts on Lys, Arg, and deoxyguanosine. As a result, systems are in place to remove these potentially damaging modifications. We have shown that the deglycase, DJ-1 , is capable of removing the intermediate aminocarbinol adduct, yielding the unmodified Arg and lactate. We are interested in fully understanding these enzymatic reactions and elucidating the consequences of protein adduction when these detoxification mechanisms are compromised. Enzymatic reactions are shown in green. https://www.galliganlab.com/publications daily 0.75 2018-08-27 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1530909796361-65J8N2MT7IO7EQY7URIF/Picture1.jpg Publications A generalized scheme of the QuARKMod platform. QuARKMod has been validated for use with a broad range of cell and tissue samples. Following protein digestion, the absolute concentrations of PTMs can be determined in a given sample. In addition, using data-dependent scanning, we outline a method for the discovery of novel or low-abundance Lys and Arg PTMs in the same biological samples. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1530911973279-0OX75HOXHJAMSNER0UGH/Picture1.jpg Publications (A) Methylglyoxal generates stable Arg protein adducts. Three isoforms of MG-H are possible with MG-H3 undergoing hydrolysis to generate CEA. Only MG-H1 and CEA are observed in physiological settings. Cells were cultured for 24 h in low-glucose medium (5 mM) and (B) cellular MGO was quantified. Chromatin was extracted from the indicated cells and subjected to QuARKMod, demonstrating basal levels of ADMA (C), MG-H1 (D), and CEA (E). Basal levels of MGO (F), chromatin ADMA (G), MG-H1 (H), and CEA (I) were also evaluated in tissues isolated from mice. Data are presented as the mean ± S.D. of three measurements. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1530911490238-627EK4NT9KL9SGLD41BK/Picture1.jpg Publications Identification of 28 site-specific MGO modifications on histones. Proteomic interrogation of site-specific MGO PTMs reveals the N-terminus of H3 and the globular domain (bold) of H2B to be heavily susceptible to modification by MGO. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526216381918-73DDX4BGX7W3PE8DQ7DB/Picture1.jpg Publications Figure 1. (A) Alkynyl-4-ONE preferentially modifies histone proteins in chromatin isolated from RKO cells. (B) Time-course of histone adduction with a4-ONE. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526239949131-TSDZEMYZI25QNSG0IJXK/Picture1.jpg Publications Application of QuARKMod for the study of in vitro enzyme activity. QuARKMod was used to investigate the overall methylation status of a synthetic peptide (2 nmol) containing a single me3Lys following incubation with a recombinant Lys demethylase, JMJD2A. A quantitative yield was observed in total methylated Lys following incubation with JMJD2A with me2Lys being the predominant PTM. The peptide sequence was as follows: ARTKQTARKme3STGGKA. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526240020290-PI4AZB8CKX8EMRYNHA0N/Picture1.jpg Publications Quantitation of Lys and Arg PTMs on isolated histones. (a) To measure histone-specific changes, histones were purified using an offline HPLC method, resulting in significant separation of each core histone. (b) Purified histones collected from HEK293 cells demonstrate varying degrees of Lys and Arg PTMs. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526216567818-F9FKBNPGBPJDQLIAFMHD/Picture1.jpg Publications Figure 2. 4-ONE adducts are located on surface-exposed amino acids. (A) Crystal structure of the nucleosome. Adducts are located at (B) H4K79, (C) H2BH82, H109 and K116, (D) H3K23 and K27 and (E) H2AH123. Structure obtained from PDB:1AOI. https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526239346858-J6AJVDGTTH51DQ02MB09/Picture1.jpg Publications Scheme 1. 4-hydroxy-2-nonenal (4-HNE) and 4-oxo-2-nonenal (4-ONE) are electrophilic molecules that adduct both DNA and protein, resulting in altered structure and function. Although similar in structure, 4-HNE and 4-ONE differ in reactivity in that 4-ONE can form stable ketoamide adducts via 1,2 addition to Lys https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1530911600122-NTBIQ7BJ9KMDOY5I8PO3/Picture1.jpg Publications DJ-1 protects chromatin from modification by MGO. (A) DJ-1 hydrolyzes the intermediate aminocarbinol product of MGO and Arg. (B) Western blotting demonstrates complete knockout of DJ-1 in both WT and GLO1-/- cell lines (DKO). (C-E) QuARKMod was performed on chromatin fractions isolated from each cohort, demonstrating significant increases in MG-H1 and CEA, and not CEL, in DKO cells compared to GLO1-/- alone. Treatments were for 6 h. Data are presented as the mean ± S.D., N = 3, and statistical significance was determined via two-way ANOVA (*** p < 0.001). https://www.galliganlab.com/join-us daily 0.75 2018-11-07 https://www.galliganlab.com/conferences daily 0.75 2018-07-17 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526506776200-RDG87OZ3GK1YPQV7WNMD/ggxwvtzz.png Conferences https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1531788794847-PWPHL16CWEOVB1IUMBX7/logo.jpg Conferences https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1526241390480-SEGNRJ8XERRRT0QYEPRL/Picture1.jpg Conferences https://www.galliganlab.com/people daily 0.75 2018-11-07 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1531521947197-DNB375ZF9BBQNTDHMI8H/Galligan+Directory.jpg People https://www.galliganlab.com/lab-news daily 0.75 2018-11-07 https://www.galliganlab.com/home daily 0.75 2018-07-27 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1531256642957-Z2VQ7O3LAF1FOEHYTT57/Picture1.jpg Home https://www.galliganlab.com/home-1 daily 1.0 2024-10-09 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1728462771499-ZEAXOLE2CW3YZV7BHRVM/unsplash-image-5A06OWU6Wuc.jpg Studying Cell Metabolism and Chromatin Function https://www.galliganlab.com/testimonials daily 0.75 2025-06-25 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/0c4b75a1-8755-4621-a8c2-0fc0dcc5fbac/Naoya.jpg The Lab - Naoya Kitamura Graduate Student - Drug Discovery and Development 2022 - Present nkitamura@pharmacy.arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1610686283178-VDHJS4N1O60NQHARPX9H/Emely+Pic.jpg The Lab - Emely Hoffman Research Technician 2020 - Present hoffmane@arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/20218377-da9d-42b7-8b77-ecfe41258e0c/Wei+Chen+Zhang.jpg The Lab - Wei Chen Zhang Graduate Student - Nutritional Sciences 2023 - Present weichenz3@arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/249ab97c-3344-4aa0-8e24-fc62613c8deb/Allison.jpg The Lab - Allison Roman Perez Undergraduate Student - Bachelor of Science in Pharmaceutical Sciences 2024 - Present aroman04@catworks.arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1628634340579-UQA8RJ2EOQEVOWWHDCGU/Jim+Headshot.jpg The Lab - James J. Galligan, PhD Principal Investigator Dr. Galligan received his PhD in Pharmacology from the University of Colorado Denver in 2012 in the laboratory of Dr. Dennis Petersen. His work was focused on the role of the endoplasmic reticulum stress and oxidative stress responses in the alcoholic liver. He then joined the laboratory of Dr. Lawrence Marnett at Vanderbilt University to complete his post-doctoral training. Here, his work elucidated two novel histone modifications stemming from endogenous electrophilic metabolites. Dr. Galligan is currently an Associate Professor at the University of Arizona in the Department of Pharmacology and Toxicology. Google Scholar Pubmed LinkedIn jgalligan@pharmacy.arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/fa561a21-1c73-4831-9192-66e67eca8d9e/Aiden.jpg The Lab - Aiden Phoebe Research Technician 2025 - Present aidenphoebe@email.arizona.edu https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1728464938607-5U8JENH8E8WSHBS19SKN/IMG_9769.jpg The Lab https://www.galliganlab.com/reserve daily 0.75 2025-05-29 https://www.galliganlab.com/methylglyoxal-metabolism-in-disease daily 0.75 2019-12-05 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1575579794639-J07N3CSG3WHL7QUJ3HKC/Methylglyoxal+Metabolism+and+Adducts+Updated.jpeg Methylglyoxal Metabolism in Disease https://www.galliganlab.com/methylglyoxal-metabolism-in-disease-1 daily 0.75 2019-01-19 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1547874930205-TBPG8P0FJ2ZK0CXZAD0O/Picture1.jpg Copy of Methylglyoxal Metabolism in Disease https://www.galliganlab.com/methylglyoxal-metabolism-in-disease-2 daily 0.75 2024-07-25 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1548306592873-U9ANCV4NL1YI4JTYT5H7/Picture1.jpg Quantifying the Post-Translational Modification Landscape https://www.galliganlab.com/contact daily 0.75 2024-01-31 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1547916290326-DPTG0UBMC5H30VNRCD2E/IMG_2943.jpg Contact https://www.galliganlab.com/methylglyoxal-metabolism-in-disease-3 daily 0.75 2019-01-25 https://images.squarespace-cdn.com/content/v1/5af5ee951aef1d1741e845af/1548290268836-14DQVY9Z7SDOYZ9GNO0W/Picture1.jpg Methylglyoxal and Chromatin Function