Dehydroalanine and dehydrobutyrine in aging and cataractous lenses reveal site-specific consequences of spontaneous protein degradation

Jessica Paredes; Zhen Wang; Purvi Patel; Kristie L Rose; Kevin L Schey

doi:10.3389/fopht.2023.1241001

Dehydroalanine and dehydrobutyrine in aging and cataractous lenses reveal site-specific consequences of spontaneous protein degradation

Front Ophthalmol (Lausanne). 2023 Oct 26:3:1241001. doi: 10.3389/fopht.2023.1241001. eCollection 2023.

Authors

Jessica Paredes¹, Zhen Wang², Purvi Patel³, Kristie L Rose^{2

3}, Kevin L Schey^{1

2

3}

Affiliations

¹ Department of Chemistry, Vanderbilt University, Nashville, TN, United States.
² Department of Biochemistry, Vanderbilt University, Nashville, TN, United States.
³ Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN, United States.

Abstract

Introduction: Protein post-translational modifications (PTMs) have been associated with aging and age-related diseases. PTMs are particularly impactful in long-lived proteins, such as those found in the ocular lens, because they accumulate with age. Two PTMs that lead to protein-protein crosslinks in aged and cataractous lenses are dehydroalanine (DHA) and dehydrobutyrine (DHB); formed from cysteine/serine and threonine residues, respectively. The purpose of this study was to quantitate DHA and DHB in human lens proteins as a function of age and cataract status.

Methods: Human lenses of various ages were divided into five donor groups: transparent lenses (18-22-year-old, 48-64-year-old, and 70-93-year-old) and cataractous human lenses of two age groups (48-64-year-old lenses, and 70-93-year-old lenses) and were subjected to proteomic analysis. Relative DHA and DHB peptide levels were quantified and compared to their non-modified peptide counterparts.

Results: For most lens proteins containing DHA or DHB, higher amounts of DHA- and DHB-modified peptides were detected in aged and cataractous lenses. DHA-containing peptides were classified into three groups based on abundance changes with age and cataract: those that (1) increased only in age-related nuclear cataract (ARNC), (2) increased in aged and cataractous lenses, and (3) decreased in aged lenses and ARNC. There was no indication that DHA or DHB levels were dependent on lens region. In most donor groups, proteins with DHA and DHB were more likely to be found among urea-insoluble proteins rather than among water- or urea-soluble proteins.

Discussion: DHA and DHB formation may induce structural effects that make proteins less soluble in water that leads to age-related protein insolubility and possibly aggregation and light scattering.

Keywords: aging; cataracts; lens; post-translational modifications; proteomics.

Abstract

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