Linked Life Data

7711043

Source:http://linkedlifedata.com/resource/pubmed/id/7711043

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
15
pubmed:dateCreated
1995-5-17
pubmed:databankReference
pubmed:abstractText
The hemolymph juvenile hormone binding protein (JHBP) from the early fifth instar larvae of Heliothis virescens (Lepidoptera, Noctuidae) has been purified, and three cDNA clones for this protein have been isolated from a fat body cDNA library constructed in bacteriophage lambda ZAP XR. The deduced amino acid sequence of the full-length clone predicts a mature protein consisting of 224 residues, a molecular mass of 24,976 Da, and a pI of 5.29. Comparison of the amino acid sequence to that of the previously described JHBP from Manduca sexta shows 51% overall identity with highly conserved N- and C-terminal regions. One of the three clones bound photoactivatable analogs of juvenile hormones with much lower affinity than the other two. This clone had Phe150 in place of the expected Cys150 conserved in other JHBP clones. The F150C mutant of this clone regained native binding affinity. For native Hvir-JHBP, the affinity for [3H]JH I was lower under reducing conditions (87 nM) relative to a 40 nM affinity under nonreducing conditions. The importance of pairs of Cys residues was addressed by preparing Cys to Ala mutants at each site. Expressed proteins were tested for binding affinity by photoaffinity labeling with tritium-labeled JH analogs and by binding assays using (10R,11S)-[3H]JH I. Curiously, the C150A mutant retained full activity, implying that the aberrant C150F was dysfunctional due to steric hindrance rather than to a missing disulfide linkage. Likewise, C29A and C194A had binding affinities unchanged from that of the full-length wild-type clone.(ABSTRACT TRUNCATED AT 250 WORDS)
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
18
pubmed:volume
34
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5234-41
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1995
pubmed:articleTitle
Key disulfide bonds in an insect hormone binding protein: cDNA cloning of a juvenile hormone binding protein of Heliothis virescens and ligand binding by native and mutant forms.
pubmed:affiliation
Department of Chemistry, State University of New York at Stony Brook 11794-3400, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S.