Linked Life Data

11241215

Source:http://linkedlifedata.com/resource/pubmed/id/11241215

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2001-3-12
pubmed:abstractText
Bone morphogenetic proteins (BMPs) belong to the transforming growth factor-beta (TGF-beta) superfamily of multifunctional cytokines. BMP induces its signal to regulate growth, differentiation, and apoptosis of various cells upon trimeric complex formation with two distinct type I and type II receptors on the cell surface: both are single-transmembrane serine/threonine kinase receptors. To identify the amino acid residues on BMP type I receptor responsible for its ligand binding, the structure-activity relationship of the extracellular ligand-binding domain of the BMP type IA receptor (sBMPR-IA) was investigated by alanine-scanning mutagenesis. The mutant receptors, as well as sBMPR-IA, were expressed as fusion proteins with thioredoxin in Escherichia coli, and purified using reverse phase high performance liquid chromatography (RP-HPLC) after digestion with enterokinase. Structural analysis of the parent protein and representative mutants in solution by CD showed no detectable differences in their folding structures. The binding affinity of the mutants to BMP-4 was determined by surface plasmon resonance biosensor. All the mutant receptors examined, with the exception of Y70A, displayed reduced affinities to BMP-4 with the rank order of decreases: I52A (17-fold) approximately F75A (15-fold) >> T64A (4-fold) = T62A (4-fold) approximately E54A (3-fold). The decreases in binding affinity observed for the latter three mutants are mainly due to decreased association rate constants while alterations in rate constants both, for association and dissociation, result in the drastically reduced affinities for the former two mutants. These results allow us to conclude that sBMPR-IA recognizes the ligand using the concave face of the molecule. The major ligand-binding site of the BMP type IA receptor consists of Phe75 in loop 2 and Ile52, Glu54, Thr62 and Thr64 on the three-stranded beta-sheet. These findings should provide a general basis for the ligand/type I receptor recognition in the TGF-beta superfamily.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
0006-3525
pubmed:author
pubmed:issnType
Print
pubmed:volume
55
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
399-406
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:11241215-Amino Acids, pubmed-meshheading:11241215-Animals, pubmed-meshheading:11241215-Binding Sites, pubmed-meshheading:11241215-Bone Morphogenetic Protein 4, pubmed-meshheading:11241215-Bone Morphogenetic Protein Receptors, Type I, pubmed-meshheading:11241215-Bone Morphogenetic Proteins, pubmed-meshheading:11241215-Circular Dichroism, pubmed-meshheading:11241215-Ligands, pubmed-meshheading:11241215-Mice, pubmed-meshheading:11241215-Models, Molecular, pubmed-meshheading:11241215-Mutagenesis, Site-Directed, pubmed-meshheading:11241215-Protein Binding, pubmed-meshheading:11241215-Protein Structure, Secondary, pubmed-meshheading:11241215-Protein Structure, Tertiary, pubmed-meshheading:11241215-Protein-Serine-Threonine Kinases, pubmed-meshheading:11241215-Receptors, Growth Factor, pubmed-meshheading:11241215-Surface Plasmon Resonance
pubmed:year
2000
pubmed:articleTitle
Identification of the ligand-binding site of the BMP type IA receptor for BMP-4.
pubmed:affiliation
Structural Biology Unit, National Institute of Agrobiological Resources, Tsukuba, Ibaraki 305-8602, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't