title: Ex vivo detection of histone H1 modified with advanced glycation end products.
abstract: A number of oxidative stress agents cause DNA and protein damage, which may compromise genomic integrity. Whereas oxidant-induced DNA damage has been extensively studied, much less is known concerning the occurrence and fate of nuclear protein damage, particularly of proteins involved in the regulation and maintenance of chromatin structure. Protein damage may be caused by the formation of reactive carbonyl species such as glyoxal, which forms after lipid peroxide degradation. It may also result from degradation of early protein glycation adducts and from methylglyoxal, formed in the process of glycolytic intermediate degradation. Major adducts indicative of protein damage include the advanced glycation end product (AGE) carboxymethyllysine (CML) and argpyrimidine protein adducts. Thus, the formation of CML and argpyrimidine protein adducts represents potential biomarkers for nuclear protein damage deriving from a variety of sources. The purpose of this study was to identify and quantify AGE adducts formed in vivo in a nuclear protein, specifically histone H1, using CML and argpyrimidine as biomarkers. Histone H1 was isolated from calf thymus collected immediately after slaughter under conditions designed to minimize AGE formation before isolation. Using antibodies directed against oxidative protein adducts, we identified CML, argpyrimidine, and protein crosslinks present in the freshly isolated histone H1. Detailed mass spectroscopy analysis of histone H1 revealed the presence of two specific lysine residues modified by CML adducts. Our results strongly suggest that glycation of important nuclear protein targets such as histone H1 occurs in vivo and that these oxidative changes may alter chromatin structure, ultimately contributing to chronic changes associated with aging and diseases such as diabetes.
authors.affiliation: Department of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57006, USA.
authors.forename: Srinath
authors.initials: S
authors.lastname: Pashikanti
authors.forename: Gilbert A
authors.initials: GA
authors.lastname: Boissonneault
authors.forename: Daniel
authors.initials: D
authors.lastname: Cervantes-Laurean
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D017127:Glycation End Products, Advanced / Q000737:chemistry / Q000378:metabolism*; D006657:Histones / Q000032:analysis* / Q000378:metabolism; D013058:Mass Spectrometry; D009687:Nuclear Proteins / Q000378:metabolism; D013950:Thymus Gland / Q000737:chemistry / Q000378:metabolism
chemical_list: D015415:Biomarkers; D017127:Glycation End Products, Advanced; D006657:Histones; D009687:Nuclear Proteins