April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Functional Analysis of Cyp1b1 Mutations in Spanish Patients With Primary Congenital Glaucoma
M.-P. Lopez-Garrido; E. Campos-Mollo; C. Blanco-Marchite; J. García-Feijoo; J. Peralta; J. Belmonte-Martínez; C. Ayuso; J. Escribano
Author Affiliations & Notes
  • M.-P. Lopez-Garrido
    Genetics, Castilla-La Mancha University Medical School, Albacete, Spain
    Cooperative Research Network on Age-Related Ocular Pathology, Visual and Life Quality, Instituto de Salud Carlos III, Madrid, Spain
  • E. Campos-Mollo
    Servicio de Oftalmología, Hospital General Universitario, Alicante, Spain
  • C. Blanco-Marchite
    Servicio de Oftalmología, Complejo Hospitalario Universitario, Albacete, Spain
  • J. García-Feijoo
    Cooperative Research Network on Age-Related Ocular Pathology, Visual and Life Quality, Instituto de Salud Carlos III, Madrid, Spain
    Servicio de Oftalmología, Hospital San Carlos, Madrid, Spain
  • J. Peralta
    Servicio de Oftalmología Infantil, Hospital La Paz, Madrid, Spain
  • J. Belmonte-Martínez
    Servicio de Oftalmología, Hospital General Universitario, Alicante, Spain
  • C. Ayuso
    Servicio de Genética, Fundación Jiménez Díaz, CIBERER, Madrid, Spain
  • J. Escribano
    Genetics, Castilla-La Mancha University Medical School, Albacete, Spain
    Cooperative Research Network on Age-Related Ocular Pathology, Visual and Life Quality, Instituto de Salud Carlos III, Madrid, Spain
  • Footnotes
    Commercial Relationships  M.-P. Lopez-Garrido, None; E. Campos-Mollo, None; C. Blanco-Marchite, None; J. García-Feijoo, None; J. Peralta, None; J. Belmonte-Martínez, None; C. Ayuso, None; J. Escribano, None.
  • Footnotes
    Support  FIS Grant PIO52494; JCCM Grant 02021-00
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 868. doi:
  • Views
  • Share
  • Tools
    • Alerts
      User Alerts
      You are adding an alert for:
      Functional Analysis of Cyp1b1 Mutations in Spanish Patients With Primary Congenital Glaucoma
      You will receive an email whenever this article is corrected, updated, or cited in the literature. You can manage this and all other alerts in My Account
      The alert will be sent to:
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation
      Citation

      M.-P. Lopez-Garrido, E. Campos-Mollo, C. Blanco-Marchite, J. García-Feijoo, J. Peralta, J. Belmonte-Martínez, C. Ayuso, J. Escribano; Functional Analysis of Cyp1b1 Mutations in Spanish Patients With Primary Congenital Glaucoma. Invest. Ophthalmol. Vis. Sci. 2009;50(13):868.

      Download citation file:

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

      ×
    • Get Permissions
  • Supplements
Abstract

Purpose: : To analyze the contribution of CYP1B1 gene mutations to primary congenital glaucoma (PCG) in Spanish patients.

Methods: : The promoter region (-1 to-867) and the 3 exons of CYP1B1 gene were analysed by PCR DNA sequencing. The CYP1B1 mutations were generated in vitro by site-directed mutagenesis using as template a cDNA encoding wild type CYP1B1 cloned into the mammalian expression vector pcDNA 3.1, and taged with the myc-epitope at the C-terminal end. To determine the enzymatic activity and protein stability of identified mutations, HEK-293T cells were transiently transfected with the corresponding cDNAs. The time course of the enzymatic activity was analyzed by determining the ethoxyresorufin O-deethylation (EROD) activityin transfected cells using a fluorimetric assay. Protein stability was studied by western blot of transfected cells exposed to cycloheximide for various time points. Recombinant CYP1B1 proteins were detected using an anti-myc antibody. Wild type CYP1B1 was used as a control in all the assays.

Results: : We found a total of 16 different mutations in 13 (34.2%) index cases. The identified mutations included 9 missense and 3 nonsense nucleotide changes, 3 small deletions and a short duplication. Eleven probands were compound heterozygotes and two were heterozygotes. Six of the identified mutations were novel. Mutations p.T404fsX30 and p.R355fsX69 were the most prevalent among index cases, and were detected in 6 (23.0%) and 3 (11.5%) patients, respectively. Functional analysis showed that 4 mutations were null-alleles while the remaining mutants were hypomorphic alleles.

Conclusions: : Our data indicate that approximately one third of Spanish patients with PCG carry loss-of-function CYP1B1 and show that null alleles are associated with the most severe phenotypes. Hypomorphic alleles may contribute to some cases of incomplete penetrance.

Keywords: mutations • genetics • enzymes/enzyme inhibitors 
© 2009, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×
This site uses cookies. By continuing to use our website, you are agreeing to our privacy policy.Accept