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Article: Interaction of collagen α1(X) containing engineered NC1 mutations with normal α1(X) in vitro. Implications for the molecular basis of Schmid metaphyseal chondrodysplasia

TitleInteraction of collagen α1(X) containing engineered NC1 mutations with normal α1(X) in vitro. Implications for the molecular basis of Schmid metaphyseal chondrodysplasia
Authors
Issue Date1999
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 1999, v. 274 n. 19, p. 13091-13097 How to Cite?
AbstractCollagen X is a short-chain homotrimeric collagen expressed in the hypertrophic zone of calcifying cartilage. The clustering of mutations in the carboxyl-terminal nonhelical NC1 domain in Schmid metaphyseal chondrodysplasia (SMCD) suggests a critical role for NC1 in collagen X structure and function. In vitro collagen X DNA expression, using T7-driven coupled transcription and translation, demonstrated that although α1(X) containing normal NC1 domains can form electrophoretically stable trimers, engineered SMCD NC1 missense or premature termination mutations prevented the formation of electrophoretically stable homotrimers or heterotrimers when co- expressed with normal α1(X). To allow the detection of more subtle interactions that may interfere with assembly but not produce SDS-stable final products, we have developed a competition-based in vitro co-expression and assembly approach. Our studies show that α1(X) chains containing SMCD mutations reduce the efficiency of normal α1(X) trimer assembly, indicating that interactions do occur between mutant and normal NC1 domains, which can impact on the formation of normal trimers. This finding has important implications for the molecular pathology of collagen X mutations in SMCD. Although we have previously demonstrated haploinsufficiency as one in vivo mechanism (Chan, D., Weng, Y. M., Hocking, A.M., Golub, S., McQuillan, D. J., and Bateman, J. F. (1998) J. Clin. Invest. 101, 1490-1499), the current study suggests dominant interference is also possible if the mutant protein is expressed in vivo. Furthermore, we establish that a conserved 13-amino acid aromatic motif (amino acids 589601) is critical for the interaction between the NC1 domains, suggesting that this region may initiate assembly and the other NC1 mutations interfered with secondary interactions important in folding or in stabilizing the assembly process.
Persistent Identifierhttp://hdl.handle.net/10722/147440
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChan, Den_US
dc.contributor.authorFreddi, Sen_US
dc.contributor.authorWeng, YMen_US
dc.contributor.authorBateman, JFen_US
dc.date.accessioned2012-05-29T06:03:44Z-
dc.date.available2012-05-29T06:03:44Z-
dc.date.issued1999en_US
dc.identifier.citationJournal Of Biological Chemistry, 1999, v. 274 n. 19, p. 13091-13097en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/147440-
dc.description.abstractCollagen X is a short-chain homotrimeric collagen expressed in the hypertrophic zone of calcifying cartilage. The clustering of mutations in the carboxyl-terminal nonhelical NC1 domain in Schmid metaphyseal chondrodysplasia (SMCD) suggests a critical role for NC1 in collagen X structure and function. In vitro collagen X DNA expression, using T7-driven coupled transcription and translation, demonstrated that although α1(X) containing normal NC1 domains can form electrophoretically stable trimers, engineered SMCD NC1 missense or premature termination mutations prevented the formation of electrophoretically stable homotrimers or heterotrimers when co- expressed with normal α1(X). To allow the detection of more subtle interactions that may interfere with assembly but not produce SDS-stable final products, we have developed a competition-based in vitro co-expression and assembly approach. Our studies show that α1(X) chains containing SMCD mutations reduce the efficiency of normal α1(X) trimer assembly, indicating that interactions do occur between mutant and normal NC1 domains, which can impact on the formation of normal trimers. This finding has important implications for the molecular pathology of collagen X mutations in SMCD. Although we have previously demonstrated haploinsufficiency as one in vivo mechanism (Chan, D., Weng, Y. M., Hocking, A.M., Golub, S., McQuillan, D. J., and Bateman, J. F. (1998) J. Clin. Invest. 101, 1490-1499), the current study suggests dominant interference is also possible if the mutant protein is expressed in vivo. Furthermore, we establish that a conserved 13-amino acid aromatic motif (amino acids 589601) is critical for the interaction between the NC1 domains, suggesting that this region may initiate assembly and the other NC1 mutations interfered with secondary interactions important in folding or in stabilizing the assembly process.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCollagen - Chemistry - Genetics - Metabolismen_US
dc.subject.meshDna Primersen_US
dc.subject.meshHumansen_US
dc.subject.meshMutationen_US
dc.subject.meshOsteochondrodysplasias - Geneticsen_US
dc.titleInteraction of collagen α1(X) containing engineered NC1 mutations with normal α1(X) in vitro. Implications for the molecular basis of Schmid metaphyseal chondrodysplasiaen_US
dc.typeArticleen_US
dc.identifier.emailChan, D:chand@hkucc.hku.hken_US
dc.identifier.authorityChan, D=rp00540en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1074/jbc.274.19.13091en_US
dc.identifier.pmid10224061-
dc.identifier.scopuseid_2-s2.0-0033531789en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0033531789&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume274en_US
dc.identifier.issue19en_US
dc.identifier.spage13091en_US
dc.identifier.epage13097en_US
dc.identifier.isiWOS:000080200400023-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridChan, D=7402216545en_US
dc.identifier.scopusauthoridFreddi, S=6602268246en_US
dc.identifier.scopusauthoridWeng, YM=7103321441en_US
dc.identifier.scopusauthoridBateman, JF=16135557700en_US

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