Abstract

Isolation of N epsilon-(gamma-glutamyl) lysine from polymers unique for cataract (Lorand et al., PNAS 78, 1356, 1981) was the spur for current efforts to investigate in greater depth the role of the protein cross-linking enzyme: transglutaminase in lens aging. The present five-year proposal represents a combination of this promising novel approach for exploring specific post-translational modifications of proteins in the aging human lens, progressing towards cataract, and in animal lens.
Specific aims i nclude the following topics: (1) selective triggering of transglutaminase- mediated cross-linking in lens--evaluation of competitive and non- competitive inhibitors; (2) N epsilon-(gamma-glutamyl) lysine in the cross-linked beta crystallin dimers; (3) identification and isolation of protein targets of transglutaminase in lens by labelling with dansylcadaverine and using anti-dansyl antibody affinity procedures--application for assessing protein associations; (4) sequencing of amino acids near the transglutaminase-reactive glutaminyl residues of beta crystallin subunits; (5) interactions of soluble proteins of the lens with dansylcadaverine-labelled beta crystallin subunits, assessed by changes in fluorescence; (6) N epsilon-(gamma-glutamyl) lysine in cross-linked polymers of human cataracts--heterogeneity, analysis of composition by immunological means and cross-link frequency; (7) N epsilon- (gamma-glutamyl) lysine content of beta crystallin dimers in aging human lens and (8) regulations of lens transglutaminase.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY003942-08
Application #
3258417
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1982-03-01
Project End
1993-02-28
Budget Start
1989-03-01
Budget End
1990-02-28
Support Year
8
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Northwestern University at Chicago
Department
Type
Schools of Arts and Sciences
DUNS #
City
Evanston
State
IL
Country
United States
Zip Code
60208

  Publications

Murthy, S N; Lomasney, J W; Mak, E C et al. (1999) Interactions of G(h)/transglutaminase with phospholipase Cdelta1 and with GTP. Proc Natl Acad Sci U S A 96:11815-9
Murthy, S N; Velasco, P T; Lorand, L (1998) Properties of purified lens transglutaminase and regulation of its transamidase/crosslinking activity by GTP. Exp Eye Res 67:273-81
Lorand, L; Stern, A M; Velasco, P T (1998) Novel inhibitors against the transglutaminase-catalysed crosslinking of lens proteins. Exp Eye Res 66:531-6
Clement, S; Velasco, P T; Murthy, S N et al. (1998) The intermediate filament protein, vimentin, in the lens is a target for cross-linking by transglutaminase. J Biol Chem 273:7604-9
Parameswaran, K N; Cheng, X F; Chen, E C et al. (1997) Hydrolysis of gamma:epsilon isopeptides by cytosolic transglutaminases and by coagulation factor XIIIa. J Biol Chem 272:10311-7
Velasco, P T; Lukas, T J; Murthy, S N et al. (1997) Hierarchy of lens proteins requiring protection against heat-induced precipitation by the alpha crystallin chaperone. Exp Eye Res 65:497-505
Clement, S; Trejo-Skalli, A V; Gu, L et al. (1997) A transglutaminase-related antigen associates with keratin filaments in some mouse epidermal cells. J Invest Dermatol 109:778-82
Lorand, L (1996) Neurodegenerative diseases and transglutaminase. Proc Natl Acad Sci U S A 93:14310-3
Trejo-Skalli, A V; Velasco, P T; Murthy, S N et al. (1995) Association of a transglutaminase-related antigen with intermediate filaments. Proc Natl Acad Sci U S A 92:8940-4
Lorand, L; Velasco, P T; Murthy, S N et al. (1992) Isolation of transglutaminase-reactive sequences from complex biological systems: a prominent lysine donor sequence in bovine lens. Proc Natl Acad Sci U S A 89:11161-3

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