March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Characterization Of Aldehyde Dehydrogenase 3A1 In Guinea Pig Corneal Extracts
Author Affiliations & Notes
  • Nicolas Crim, Sr.
    Departament of Ophthalmology, University Clinic Reina Fabiola, Universidad Católica de Córdoba, Argentina
  • María F. Suarez
    Departament of Clinical Biochemistry, CIBICI, Faculty of Chemical Sciences, Universidad Nacional de Cordoba, Argentina
  • María E. Forniés Paz
    Departament of Ophthalmology, University Clinic Reina Fabiola, Universidad Católica de Córdoba, Argentina
  • Rodolfo Monti
    Departament of Ophthalmology, University Clinic Reina Fabiola, Universidad Católica de Córdoba, Argentina
  • Evangelina Espósito
    Departament of Ophthalmology, University Clinic Reina Fabiola, Universidad Católica de Córdoba, Argentina
  • Julio A. Urrets-Zavalía
    Departament of Ophthalmology, University Clinic Reina Fabiola, Universidad Católica de Córdoba, Argentina
  • Thamara A. Cafaro
    Departament of Clinical Biochemistry, CIBICI, Faculty of Chemical Sciences, Universidad Nacional de Cordoba, Argentina
  • Horacio M. Serra
    Departament of Clinical Biochemistry, CIBICI, Faculty of Chemical Sciences, Universidad Nacional de Cordoba, Argentina
  • Footnotes
    Commercial Relationships  Nicolas Crim, Sr., None; María F. Suarez, None; María E. Forniés Paz, None; Rodolfo Monti, None; Evangelina Espósito, None; Julio A. Urrets-Zavalía, None; Thamara A. Cafaro, None; Horacio M. Serra, None
  • Footnotes
    Support  Supported by CONICET (PIP 112-200801-01455), and FONCYT (PICT 38052)
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 4226. doi:
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      Nicolas Crim, Sr., María F. Suarez, María E. Forniés Paz, Rodolfo Monti, Evangelina Espósito, Julio A. Urrets-Zavalía, Thamara A. Cafaro, Horacio M. Serra; Characterization Of Aldehyde Dehydrogenase 3A1 In Guinea Pig Corneal Extracts. Invest. Ophthalmol. Vis. Sci. 2012;53(14):4226.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Aldehyde dehydrogenase 3A1 (ALDH3A1) is a corneal water-soluble protein described in many mammals with an important role in the protection of ocular structures against oxidative damage. Since this enzyme has not been characterized in guinea pig (GP) we investigated expression, molecular aspects and functionality of ALDH3A1 in GP corneas.

Methods: : Epithelial and stromal corneal extracts from 10 healthy 3-5 months-old male albinos GP from Ssi:AL strain were used. Animals were used adhering to ARVO statement for animals in vision research. The expression of ALDH3A1 was studied by means of Immunoblotting. The activity and specificity toward its substrate (hexanal or benzaldehyde) was studied by spectrophotometry and zymography assays, and the existence of isoforms by isoelectric focusing.

Results: : ALDH3A1 was only identified in GP corneal epithelium. The enzyme has an apparent molecular weight of 54 KDa. Using NADP+ as a coenzyme, it was observed that ALDH3A1 has higher affinity for benzaldehyde than hexanal (Km: 45.15 vs 98.40), and the enzymatic activity was 0.0035+/-0.0026 UI/mg of proteins vs 0.0024+/-0.0072 UI/mg.Isoelectric focusing followed by enzyme staining in the gel revealed two corneal isozyme species, migrating at a pH 10.

Conclusions: : For the first time, ALDH3A1 has been identified and described in the albino GP corneal epithelium using different methods. ALDH3A1 from these animals presents some unique characteristics compared with other mammals.

Keywords: cornea: epithelium • protein purification and characterization • protective mechanisms 
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