title: Identification of Sequence Similarities among Isomerization Hotspots in Crystallin Proteins.
abstract: The eye lens crystallins represent an ideal target for studying the effects of aging on protein structure. Herein we examine separately the water-soluble (WS) and water-insoluble (WI) crystallin fractions and identify sites of isomerization and epimerization. Both collision-induced dissociation and radical-directed dissociation are needed for detection of these non-mass-shifting post-translational modifications. Isomerization levels differ significantly between the WS and the WI fractions from sheep, pig, and cow eye lenses. Residues that are most susceptible to isomerization are identified site-specifically and are found to reside in structurally disordered regions. However, isomerization in structured domains, although less common, often yields more dramatic effects on solubility. Numerous isomerization hotspots were also identified and occur in regions with aspartic acid and serine repeats. For example,128KMEIVDDDVPSLW140in βB3 crystallin contains three sequential aspartic acid residues and is isomerized heavily in the WI fractions, while it is not modified at all in the WS fractions. Potential causes for enhanced isomerization at sites with acidic residue repeats are presented. The importance of acidic residue repeats extends beyond the lens, as they are found in many other long-lived proteins associated with disease.
authors.affiliation: Department of Chemistry, University of California , Riverside, 501 Big Springs Road, Riverside, California 92521, United States.
authors.forename: Yana A
authors.initials: YA
authors.lastname: Lyon
authors.affiliation: Department of Chemistry, University of California , Riverside, 501 Big Springs Road, Riverside, California 92521, United States.
authors.forename: Georgette M
authors.initials: GM
authors.lastname: Sabbah
authors.affiliation: Department of Chemistry, University of California , Riverside, 501 Big Springs Road, Riverside, California 92521, United States.
authors.forename: Ryan R
authors.identifier: 0000-0003-1580-8355
authors.initials: RR
authors.lastname: Julian
mesh_terms: D000375:Aging / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D002386:Cataract / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002417:Cattle; D003459:Crystallins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000235:genetics; D012756:Sheep; D012995:Solubility; D013552:Swine
chemical_list: D003459:Crystallins
keywords: epimerization; isomerization; long-lived proteins; photodissociation