| Understanding carbamoyl-phosphate synthetase I (CPS1) deficiency by using expression studies and structure-based analysis. | |
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MedLine Citation:
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PMID: 20578160 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Carbamoyl-phosphate synthetase I (CPS1) deficiency (CPS1D), a recessively inherited urea cycle error due to CPS1 gene mutations, causes life-threatening hyperammonemia. The disease-causing potential of missense mutations in CPS1 deficiency can be ascertained with the recombinant CPS1 expression and purification system reported here, which uses baculovirus and insect cells. We study with this system the effects of nine clinical mutations and one polymorphism on CPS1 solubility, stability, activity, and kinetic parameters for NAG. Five of the mutations (p.T471N, p.Q678P, p.P774L, p.R1453Q, and p.R1453W) are first reported here, in three severe CPS1D patients. p.P774L, p.R1453Q, and p.R1453W inactivate CPS1, p.T471N and p.Y1491H greatly decrease the apparent affinity for NAG, p.Q678P hampers correct enzyme folding, and p.S123F, p.H337R, and p.P1411L modestly decrease activity. p.G1376S is confirmed a trivial polymorphism. The effects of the C-terminal domain mutations are rationalized in the light of this domain crystal structure, including the NAG site structure [Pekkala et al. Biochem J 424:211-220]. The agreement of clinical observations and in vitro findings, and the possibility to identify CPS1D patients who might benefit from specific treatment with NAG analogues because they exhibit reduced affinity for NAG highlight the value of this novel CPS1 expression/purification system. |
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Authors:
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Satu Pekkala; Ana I Martínez; Belén Barcelona; Igor Yefimenko; Ulrich Finckh; Vicente Rubio; Javier Cervera |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Human mutation Volume: 31 ISSN: 1098-1004 ISO Abbreviation: Hum. Mutat. Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-07-01 Completed Date: 2010-10-04 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9215429 Medline TA: Hum Mutat Country: United States |
Other Details:
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Languages: eng Pagination: 801-8 Citation Subset: IM |
Copyright Information:
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(c) 2010 Wiley-Liss, Inc. |
Affiliation:
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Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Binding Sites / genetics Carbamoyl-Phosphate Synthase (Ammonia) / chemistry, genetics*, metabolism Carbamoyl-Phosphate Synthase I Deficiency Disease / enzymology, genetics* Cell Line Enzyme Stability Glutamine / analogs & derivatives, metabolism Humans Kinetics Models, Molecular Mutagenesis, Site-Directed Mutant Proteins / genetics, metabolism Mutation* Polymorphism, Genetic* Protein Folding Protein Structure, Tertiary Rats Recombinant Proteins / chemistry, metabolism Spodoptera Structure-Activity Relationship Substrate Specificity Temperature |
| Chemical | |
Reg. No./Substance:
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0/Mutant Proteins; 0/Recombinant Proteins; 2490-97-3/aceglutamide; 56-85-9/Glutamine; EC 6.3.4.16/Carbamoyl-Phosphate Synthase (Ammonia) |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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