Linked Life Data

7548010

Source:http://linkedlifedata.com/resource/pubmed/id/7548010

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
39
pubmed:dateCreated
1995-11-8
pubmed:abstractText
The structure of the ca. 250 amino acid hormone binding domain of the human estrogen receptor (hER-LBD), expressed in E. coli and purified as a complex with estradiol, has been probed by selective proteolysis, with analysis of the protein fragments both by classical methods (SDS-PAGE and Edman N-terminal sequencing) and by mass spectrometry (HPLC-coupled electrospray ionization mass spectrometry (LC/ESI-MS)). Rapid cleavage by several proteases (trypsin, chymotrypsin, thermolysin, and Asp-N endoproteinase) is observed within a localized region (residues 297-303) at the N-terminus. In contrast, proteolytic scission at the C-terminus is less localized and more progressive; initial cuts by trypsin, chymotrypsin, thermolysin, V8, and Asp-N proteinases are observed to occur in the region 553-571, followed by further cleavage with thermolysin (548) and trypsin (548, 531, and 529). Thus, N304 and K529 define the protease-resistant N- and C-termini of a core structure for this domain that appears to contain the elements sufficient for ligand binding. The remaining segment of this domain (530-553), which is known to embody elements essential for ligand-modulated transcription activation (AF-2), is likely a surface-exposed region that, through these studies, is shown to be accessible to proteases. Only a single region within the 26 kDa ligand-binding core (N304-K529) has been identified as being readily accessible to proteases; rapid proteolysis using the proteases trypsin, chymotrypsin, and thermolysin, is localized to residues 465-468, with cleavage occurring at residues K467, L466, and both T465 and S468, respectively. The flexibility implied by the cuts in this internal 465-468 region suggest that the hER-LBD may actually consist of two subdomains. These proteolysis studies provide a substantially refined view of the conformational nature of the human estrogen receptor ligand binding domain.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
3
pubmed:volume
34
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
12605-15
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:7548010-Amino Acid Sequence, pubmed-meshheading:7548010-Binding Sites, pubmed-meshheading:7548010-Chromatography, High Pressure Liquid, pubmed-meshheading:7548010-Chymotrypsin, pubmed-meshheading:7548010-Endopeptidases, pubmed-meshheading:7548010-Estradiol, pubmed-meshheading:7548010-Humans, pubmed-meshheading:7548010-Hydrolysis, pubmed-meshheading:7548010-Ligands, pubmed-meshheading:7548010-Mass Spectrometry, pubmed-meshheading:7548010-Metalloendopeptidases, pubmed-meshheading:7548010-Molecular Sequence Data, pubmed-meshheading:7548010-Peptide Mapping, pubmed-meshheading:7548010-Receptors, Estrogen, pubmed-meshheading:7548010-Sequence Homology, Amino Acid, pubmed-meshheading:7548010-Serine Endopeptidases, pubmed-meshheading:7548010-Thermolysin, pubmed-meshheading:7548010-Trypsin
pubmed:year
1995
pubmed:articleTitle
Analysis of the structural core of the human estrogen receptor ligand binding domain by selective proteolysis/mass spectrometric analysis.
pubmed:affiliation
Department of Chemistry, University of Illinois, Urbana 61801, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't