Age-dependent Deamidation of Lifelong Proteins in the Human Lens

Overview

Journal Invest Ophthalmol Vis Sci

Specialty Ophthalmology

Date 2010 Jan 8

PMID 20053973

Citations 69

Authors

Peter G Hains

Roger J W Truscott

Affiliations

Soon will be listed here.

Abstract

Purpose: Deamidation is a common posttranslational modification in human lens crystallins and may be a key factor in the age-related denaturation of such lifelong proteins. The aim of this study was to identify the sites of deamidation in older lenses.

Methods: High-performance liquid chromatography/mass spectrometry of tryptic digests was used to identify sites of deamidation in the major human lens crystallins. Older normal and age-matched cataractous lenses were compared with fetal lenses.

Results: Approximately equal numbers of glutamine and asparagine residues were deamidated in older lenses; however, the extent of deamidation of Asn was three times greater than that of Gln (Asn, 22.6% +/- 3.6%; Gln, 6.6% +/- 1.3%). Individual crystallins differed markedly in their extent of deamidation, and deamidated residues were typically localized within discrete regions of the polypeptides. A large percentage (42%) of the sites of deamidation were characterized by the presence of a basic amino acid one residue removed from the original Gln or Asn. At nine such sites, the extent of Asn deamidation averaged 50% in aged lenses. There were few differences in deamidation between crystallins of aged normal and nuclear cataractous lenses.

Conclusions: Equal numbers of Asn and Gln residues are deamidated in crystallins from aged normal and cataractous lenses. Deamidation of Asn/Gln in lifelong proteins, such as those in the lens, may be governed to a significant degree by base-catalyzed processes.

Citing Articles

Enhancing the Thermostability of Alkaline Protease 2709 by Computation-Based Rational Design.

Yuan Y, Zhao G, Lu J, Wang L, Shi Y, Zhang J Molecules. 2025; 30(5).

PMID: 40076384 PMC: 11901772. DOI: 10.3390/molecules30051160.

Cumulative asparagine to aspartate deamidation fails to perturb γD-crystallin structure and stability.

Guseman A, Gonzalez J, Yang D, Gronenborn A Protein Sci. 2024; 33(8):e5120.

PMID: 39022918 PMC: 11255865. DOI: 10.1002/pro.5120.

Lens Cytoskeleton: An Update on the Etiopathogenesis of Human Cataracts.

Karakosta C, Samiotaki M, Panayotou G, Papaconstantinou D, Moschos M Cureus. 2024; 16(3):e56793.

PMID: 38650819 PMC: 11035009. DOI: 10.7759/cureus.56793.

HLA-DQB1*05 subtypes and not DRB1*10:01 mediates risk in anti-IgLON5 disease.

Yogeshwar S, Muniz-Castrillo S, Sabater L, Peris-Sempere V, Mallajosyula V, Luo G Brain. 2024; 147(7):2579-2592.

PMID: 38425314 PMC: 11224611. DOI: 10.1093/brain/awae048.

Eye lens β-crystallins are predicted by native ion mobility-mass spectrometry and computations to form compact higher-ordered heterooligomers.

Rolland A, Takata T, Donor M, Lampi K, Prell J Structure. 2023; 31(9):1052-1064.e3.

PMID: 37453416 PMC: 10528727. DOI: 10.1016/j.str.2023.06.013.

References

Basak A, Bateman O, Slingsby C, Pande A, Asherie N, Ogun O . High-resolution X-ray crystal structures of human gammaD crystallin (1.25 A) and the R58H mutant (1.15 A) associated with aculeiform cataract. J Mol Biol. 2003; 328(5):1137-47. DOI: 10.1016/s0022-2836(03)00375-9. View

Robinson N, Robinson A . Deamidation of human proteins. Proc Natl Acad Sci U S A. 2001; 98(22):12409-13. PMC: 60067. DOI: 10.1073/pnas.221463198. View

Wilmarth P, Tanner S, Dasari S, Nagalla S, Riviere M, Bafna V . Age-related changes in human crystallins determined from comparative analysis of post-translational modifications in young and aged lens: does deamidation contribute to crystallin insolubility?. J Proteome Res. 2006; 5(10):2554-66. PMC: 2536618. DOI: 10.1021/pr050473a. View

Kim Y, Kapfer D, Boekhorst J, Lubsen N, Bachinger H, Shearer T . Deamidation, but not truncation, decreases the urea stability of a lens structural protein, betaB1-crystallin. Biochemistry. 2002; 41(47):14076-84. DOI: 10.1021/bi026288h. View

Takemoto L, Boyle D . Increased deamidation of asparagine during human senile cataractogenesis. Mol Vis. 2000; 6:164-8. View

Geiger T, Clarke S . Deamidation, isomerization, and racemization at asparaginyl and aspartyl residues in peptides. Succinimide-linked reactions that contribute to protein degradation. J Biol Chem. 1987; 262(2):785-94. View

Lampi K, Ma Z, Shih M, Shearer T, Smith J, Smith D . Sequence analysis of betaA3, betaB3, and betaA4 crystallins completes the identification of the major proteins in young human lens. J Biol Chem. 1997; 272(4):2268-75. DOI: 10.1074/jbc.272.4.2268. View

Dasari S, Wilmarth P, Rustvold D, Riviere M, Nagalla S, David L . Reliable detection of deamidated peptides from lens crystallin proteins using changes in reversed-phase elution times and parent ion masses. J Proteome Res. 2007; 6(9):3819-26. DOI: 10.1021/pr070182x. View

Takemoto L, Boyle D . Specific glutamine and asparagine residues of gamma-S crystallin are resistant to in vivo deamidation. J Biol Chem. 2000; 275(34):26109-12. DOI: 10.1074/jbc.M002809200. View

10.

Zhang Z, David L, Smith D, Smith J . Resistance of human betaB2-crystallin to in vivo modification. Exp Eye Res. 2001; 73(2):203-11. DOI: 10.1006/exer.2001.1023. View

11.

Lapko V, Purkiss A, Smith D, Smith J . Deamidation in human gamma S-crystallin from cataractous lenses is influenced by surface exposure. Biochemistry. 2002; 41(27):8638-48. DOI: 10.1021/bi015924t. View

12.

Coghlan S, Augusteyn R . Changes in the distribution of proteins in the aging human lens. Exp Eye Res. 1977; 25(6):603-11. DOI: 10.1016/0014-4835(77)90139-7. View

13.

Sathish H, Koteiche H, Mchaourab H . Binding of destabilized betaB2-crystallin mutants to alpha-crystallin: the role of a folding intermediate. J Biol Chem. 2004; 279(16):16425-32. DOI: 10.1074/jbc.M313402200. View

14.

Padgaonkar V, Giblin F, Reddy V . Disulfide cross-linking of urea-insoluble proteins in rabbit lenses treated with hyperbaric oxygen. Exp Eye Res. 1989; 49(5):887-99. DOI: 10.1016/s0014-4835(89)80047-8. View

15.

Shapira R, Wilkinson K, Shapira G . Racemization of individual aspartate residues in human myelin basic protein. J Neurochem. 1988; 50(2):649-54. DOI: 10.1111/j.1471-4159.1988.tb02960.x. View

16.

Flaugh S, Mills I, King J . Glutamine deamidation destabilizes human gammaD-crystallin and lowers the kinetic barrier to unfolding. J Biol Chem. 2006; 281(41):30782-93. DOI: 10.1074/jbc.M603882200. View

17.

Takata T, Oxford J, Brandon T, Lampi K . Deamidation alters the structure and decreases the stability of human lens betaA3-crystallin. Biochemistry. 2007; 46(30):8861-71. PMC: 2597435. DOI: 10.1021/bi700487q. View

18.

Truscott R . Age-related nuclear cataract-oxidation is the key. Exp Eye Res. 2005; 80(5):709-25. DOI: 10.1016/j.exer.2004.12.007. View

19.

Takemoto L, Boyle D . Deamidation of specific glutamine residues from alpha-A crystallin during aging of the human lens. Biochemistry. 1998; 37(39):13681-5. DOI: 10.1021/bi981542k. View

20.

Robinson N, Robinson A, MERRIFIELD R . Mass spectrometric evaluation of synthetic peptides as primary structure models for peptide and protein deamidation. J Pept Res. 2001; 57(6):483-93. DOI: 10.1034/j.1399-3011.2001.00863.x. View