|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 275, Issue 46, 36415-36422, November 17, 2000
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Medical Research Council Laboratory for Molecular Cell
Biology and the Department of Biology, University College London,
Gower Street, London WC1E 6BT, United Kingdom
The autosomal recessive disorder primary
hyperoxaluria type 1 (PH1) is caused by a deficiency of the
liver-specific pyridoxal-phosphate-dependent enzyme
alanine:glyoxylate aminotransferase (AGT). Numerous mutations and
polymorphisms in the gene encoding AGT have been identified, but in
only a few cases has the causal relationship between genotype and
phenotype actually been demonstrated. In this study, we have determined
the effects of the most common naturally occurring amino acid
substitutions (both normal polymorphisms and disease-causing mutations)
on the properties, especially specific catalytic activity, of purified
recombinant AGT. The results presented in this paper show the
following: 1) normal human His-tagged AGT can be expressed at high
levels in Escherichia coli and purified in a correctly folded, dimerized and catalytically active state; 2) presence of the
common P11L polymorphism decreases the specific activity of purified
recombinant AGT by a factor of three; 3) AGTs containing four of the
most common PH1-specific mutations (G41R, F152I, G170R, and I244T) are
all soluble and catalytically active in the absence of the P11L
polymorphism, but in its presence all lead to protein destabilization
and aggregation into inclusion bodies; 4) naturally occurring and
artificial amino acid substitutions that lead to peroxisome-to-mitochondrion AGT mistargeting in mammalian cells also
lead to destabilization and aggregation in E. coli; and 5) the PH1-specific G82E mutation abolishes AGT catalytic activity by
interfering with cofactor binding, as does the artificial K209R mutation at the putative site of cofactor Shiff base formation. These
results are discussed in the light of the high allelic frequency (~20%) of the P11L polymorphism and its importance in determining the phenotypic manifestations of mutations in PH1.
Functional Synergism between the Most Common Polymorphism in
Human Alanine:Glyoxylate Aminotransferase and Four of the Most Common
Disease-causing Mutations*
*
This work was supported by the Medical Research Council.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: MRC Laboratory for
Molecular Cell Biology, University College London, Gower St., London
WC1E 6BT, UK. E-mail: c.danpure@ ucl.ac.uk.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  B. Cellini, R. Montioli, A. Paiardini, A. Lorenzetto, and C. B. Voltattorni Molecular Insight into the Synergism between the Minor Allele of Human Liver Peroxisomal Alanine:Glyoxylate Aminotransferase and the F152I Mutation J. Biol. Chem., March 27, 2009; 284(13): 8349 - 8358. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. D. Hopper, A. M. C. Pittman, M. C. Fitzgerald, and C. L. Tucker In Vivo and in Vitro Examination of Stability of Primary Hyperoxaluria-associated Human Alanine:Glyoxylate Aminotransferase J. Biol. Chem., November 7, 2008; 283(45): 30493 - 30502. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Williams and G. Rumsby Selected Exonic Sequencing of the AGXT Gene Provides a Genetic Diagnosis in 50% of Patients with Primary Hyperoxaluria Type 1 Clin. Chem., July 1, 2007; 53(7): 1216 - 1221. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. G. Monico, S. Rossetti, H. A. Schwanz, J. B. Olson, P. A. Lundquist, D. B. Dawson, P. C. Harris, and D. S. Milliner Comprehensive Mutation Screening in 55 Probands with Type 1 Primary Hyperoxaluria Shows Feasibility of a Gene-Based Diagnosis J. Am. Soc. Nephrol., June 1, 2007; 18(6): 1905 - 1914. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. C. Salido, X. M. Li, Y. Lu, X. Wang, A. Santana, N. Roy-Chowdhury, A. Torres, L. J. Shapiro, and J. Roy-Chowdhury Alanine-glyoxylate aminotransferase-deficient mice, a model for primary hyperoxaluria that responds to adenoviral gene transfer PNAS, November 28, 2006; 103(48): 18249 - 18254. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. J. Danpure Primary hyperoxaluria: from gene defects to designer drugs? Nephrol. Dial. Transplant., August 1, 2005; 20(8): 1525 - 1529. [Full Text] [PDF]
|
|
|
|
|
|
P. A. J. Huber, G. M. Birdsey, M. J. Lumb, D. T. R. Prowse, T. J. Perkins, D. R. Knight, and C. J. Danpure Peroxisomal Import of Human Alanine:glyoxylate Aminotransferase Requires Ancillary Targeting Information Remote from Its C Terminus J. Biol. Chem., July 22, 2005; 280(29): 27111 - 27120. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Santana, E. Salido, A. Torres, and L. J. Shapiro Primary hyperoxaluria type 1 in the Canary Islands: A conformational disease due to I244T mutation in the P11L-containing alanine:glyoxylate aminotransferase PNAS, June 10, 2003; 100(12): 7277 - 7282. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Holbrook and C. J. Danpure Molecular Basis for the Dual Mitochondrial and Cytosolic Localization of Alanine:Glyoxylate Aminotransferase in Amphibian Liver Cells J. Biol. Chem., January 11, 2002; 277(3): 2336 - 2344. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |