Linked Life Data

1694853

Source:http://linkedlifedata.com/resource/pubmed/id/1694853

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
20
pubmed:dateCreated
1990-8-14
pubmed:databankReference
pubmed:abstractText
A lambda gt11 expression library containing cDNA from total chick embryo was screened with S103L, a rat monoclonal antibody which reacts specifically with the core protein of the chick cartilage chondroitin sulfate proteoglycan. One clone was identified which produced a 220-kDa beta-galactosidase/S103L-binding fusion protein. Sequencing the entire 1.5-kilobase cDNA insert showed that it contained a single open reading frame, which encoded a portion of the proteoglycan core protein from the chondroitin sulfate domain. This was confirmed by comparison with amino acid sequence data from peptide CS-B, which was derived from the chondroitin sulfate domain (Krueger, R.C., Jr., Fields, T. A., Hildreth, J., IV, and Schwartz, N.B. (1990) J. Biol. Chem. 265, 12075-12087). Furthermore, the 3' end of the insert overlapped with 23 bases at the 5' end of the published sequence for the C-terminal globular domain (Sai, S., Tanaka, T., Kosher, R. A., and Tanzer, M. L. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 5081-5085), which oriented this clone, as well as the CS peptide, along the protein core. The cDNA insert hybridized with a 9-kilobase mRNA from sternal chondrocytes as well as a similar sized message in brain but did not hybridize to any message from rat chondrosarcoma or from undifferentiated limb bud mesenchyme. In further studies, the fusion protein as well as a cyanogen bromide fragment (70 kDa) derived from it were isolated and shown to react with S103L, indicating that cleavage at methionine residues does not disrupt the antibody recognition site. Purification and N-terminal sequencing of the antigenic CNBr fragment derived from the fusion protein revealed that its N terminus is preceded by a methionine in the fusion protein and overlaps with the N terminus of peptide CS-B. As peptide CS-B is not recognized by S103L and the C terminus of peptide CS-B lies beyond the proteoglycan portion of the antigenic CNBr fragment, the S103L epitope is either contained within the 11 amino acids preceding the N terminus of peptide CS-B or it spans the clostripain cleavage site at the origin of the N terminus of peptide CS-B.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
265
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
12088-97
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:1694853-Aggrecans, pubmed-meshheading:1694853-Amino Acid Sequence, pubmed-meshheading:1694853-Animals, pubmed-meshheading:1694853-Base Sequence, pubmed-meshheading:1694853-Cartilage, pubmed-meshheading:1694853-Chick Embryo, pubmed-meshheading:1694853-Chickens, pubmed-meshheading:1694853-Chondroitin Sulfate Proteoglycans, pubmed-meshheading:1694853-Cloning, Molecular, pubmed-meshheading:1694853-Cyanogen Bromide, pubmed-meshheading:1694853-DNA, pubmed-meshheading:1694853-Epitopes, pubmed-meshheading:1694853-Extracellular Matrix Proteins, pubmed-meshheading:1694853-Gene Library, pubmed-meshheading:1694853-Glycoproteins, pubmed-meshheading:1694853-Lectins, C-Type, pubmed-meshheading:1694853-Models, Structural, pubmed-meshheading:1694853-Molecular Sequence Data, pubmed-meshheading:1694853-Peptide Mapping, pubmed-meshheading:1694853-Proteoglycans, pubmed-meshheading:1694853-Recombinant Fusion Proteins, pubmed-meshheading:1694853-Restriction Mapping, pubmed-meshheading:1694853-Sequence Homology, Nucleic Acid
pubmed:year
1990
pubmed:articleTitle
Chick cartilage chondroitin sulfate proteoglycan core protein. II. Nucleotide sequence of cDNA clone and localization of the S103L epitope.
pubmed:affiliation
Department of Pediatrics, University of Chicago, Illinois 60637.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S.