Linked Life Data

9565588

Source:http://linkedlifedata.com/resource/pubmed/id/9565588

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
19
pubmed:dateCreated
1998-6-5
pubmed:abstractText
Heregulins (HRGs) are epidermal growth factor (egf) domain containing polypeptide growth factors that bind and activate several members of the ErbB receptor family. Although HRG can bind to ErbB3 and ErbB4 homodimers, the highest affinity and most intracellularly active receptor complexes are hetero-oligomers containing ErbB2. The HRGbeta egf domain was displayed on the surface of M13 phage to facilitate mutagenic analysis and optimize for binding to a homodimeric ErbB3-immunoglobulin (IgG) fusion. Nine libraries were constructed in which virtually the entire sequence was randomized in stretches of four to six amino acids. These were selected separately for binding to immobilized ErbB3-IgG. Analysis of the resulting sequences revealed some areas that diverged radically from the wild-type, whereas others showed strong conservation. The degree of wild-type conservation correlated strongly with the functional importance of the residues as determined by alanine scanning mutagenesis (Jones, J. T., Ballinger, M. D., Pisacane, P. I., Lofgren, J. A., Fitzpatrick, V. D., Fairbrother, W. J., Wells, J. A., and Sliwkowski, M. X. (1998) J. Biol. Chem. 273, 11667-11674). Some variants from several libraries showed significant improvements in binding affinity to the ErbB3-IgG. These optimized segments were combined in various ways in the same molecule to generate variants (containing up to 16 mutations) that had >50-fold higher affinity than wild-type HRGbeta. The optimized variants stimulated ErbB2 phophorylation on MCF7 cells at levels similar to wild-type. This indicates wild-type affinity is optimized for potency and that factors other than affinity for ErbB3 are limiting. These variants showed enhanced affinity toward the ErbB4 homodimer, suggesting these receptors use very similar binding determinants despite them having 65% sequence identity.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
8
pubmed:volume
273
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
11675-84
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:9565588-Amino Acid Sequence, pubmed-meshheading:9565588-Bacteriophage M13, pubmed-meshheading:9565588-Binding Sites, pubmed-meshheading:9565588-Carrier Proteins, pubmed-meshheading:9565588-Glycoproteins, pubmed-meshheading:9565588-Ligands, pubmed-meshheading:9565588-Models, Molecular, pubmed-meshheading:9565588-Molecular Sequence Data, pubmed-meshheading:9565588-Neuregulin-1, pubmed-meshheading:9565588-Nuclear Magnetic Resonance, Biomolecular, pubmed-meshheading:9565588-Peptide Library, pubmed-meshheading:9565588-Protein Binding, pubmed-meshheading:9565588-Proto-Oncogene Proteins, pubmed-meshheading:9565588-Receptor, Epidermal Growth Factor, pubmed-meshheading:9565588-Receptor, erbB-3, pubmed-meshheading:9565588-Recombinant Proteins, pubmed-meshheading:9565588-Structure-Activity Relationship
pubmed:year
1998
pubmed:articleTitle
Selection of heregulin variants having higher affinity for the ErbB3 receptor by monovalent phage display.
pubmed:affiliation
Department of Protein Engineering, Genentech, Incorporated, South San Francisco, California 94080, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.