Linked Life Data

20809901

Source:http://linkedlifedata.com/resource/pubmed/id/20809901

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2010-11-15
pubmed:abstractText
Three mutations of the B4GALT7 gene [encoding ?1,4-GalT7 (?1,4-galactosyltransferase 7)], corresponding to A186D, L206P and R270C, have been identified in patients with the progeroid form of the Ehlers-Danlos syndrome and are described as being associated with the reduction or loss of ?1,4-GalT7 activity. However, the molecular basis of the reduction or loss of activity remained to be determined. In the present study, wild-type, A186D, L206P and R270C ?1,4-GalT7 were expressed in CHO618 cells as membrane proteins and in Escherichia coli as soluble proteins fused to MBP (maltose-binding protein). The ability of the expressed proteins to transfer galactose from donor to acceptor substrates was systematically characterized by kinetic analysis. The physicochemical properties of soluble proteins were explored by isothermal titration calorimetry, which is a method of choice when determining the thermodynamic parameters of the binding of substrates. Together, the results showed that: (i) the L206P mutation abolished the activity when L206P ?1,4GalT7 was either inserted in the membrane or expressed as a soluble MBP-full-length fusion protein; (ii) the A186D mutation weakly impaired the binding of the donor substrate; and (iii) the R270C mutation strongly impaired the binding of the acceptor substrate. Moreover, the ex vivo consequences of the mutations were investigated by evaluating the priming efficiency of xylosides on GAG (glycosaminoglycan) chain initiation. The results demonstrate a quantitative effect on GAG biosynthesis, depending on the mutation; GAG biosynthesis was fully inhibited by the L206P mutation and decreased by the R270C mutation, whereas the A186D mutation did not affect GAG biosynthesis severely.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1470-8728
pubmed:author
pubmed:issnType
Electronic
pubmed:day
1
pubmed:volume
432
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
303-11
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Biochemical and thermodynamic characterization of mutated ?1,4-galactosyltransferase 7 involved in the progeroid form of the Ehlers-Danlos syndrome.
pubmed:affiliation
ARN-RNP-structure fonction-maturation, Enzymologie Moléculaire et Structurale, Unité Mixte de Recherche 7214, CNRS-Nancy Université, Faculté des Sciences et Techniques, BP 239 Vandoeuvre-Les-Nancy, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't