Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
31
|
| pubmed:dateCreated |
1991-12-13
|
| pubmed:abstractText |
A portion of rat mannose-binding protein A (MBP-A), a Ca(2+)-dependent animal lectin, has been overproduced in a bacterial expression system, biochemically characterized, and crystallized. A fragment corresponding to the COOH-terminal 115 residues of native MBP-A, produced by subtilisin digestion of the bacterially expressed protein, contains the carbohydrate-recognition domain (CRD). Gel filtration, chemical cross-linking, and crystallographic self-rotation function analyses indicate that the subtilisin fragment is a dimer, although the complete bacterially expressed fragment, containing the neck and CRD of MBP-A, is a trimer. Crystals of the minimal CRD, obtained only as a complex with a Man6GlcNAc2Asn glycopeptide, diffract to Bragg spacings of at least 1.7 A. Several trivalent lanthanide ions (Ln3+) can substitute for Ca2+, as assessed by their ability to support carbohydrate binding and to protect the CRD from proteolysis in a manner similar to that observed for Ca2+. These assays indicate that Ln2+ binds about 30 times more tightly than Ca2+ to the CRD, and that two Ca2+ or Ln3+ bind to each monomer, a result confirmed by determination of the Ho3+ positions in a Ho(3+)-containing crystal of the CRD. Crystals grown in the presence of Ln3+ belong to different space groups from those obtained with Ca2+ and are therefore not useable for traditional crystallographic phase determination methods, but are well-suited for high resolution structure determination by multiwavelength anomalous dispersion phasing.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Cations,
http://linkedlifedata.com/resource/pubmed/chemical/Lanthanum,
http://linkedlifedata.com/resource/pubmed/chemical/Lectins,
http://linkedlifedata.com/resource/pubmed/chemical/Mannose-Binding Lectins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0021-9258
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
5
|
| pubmed:volume |
266
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
20678-86
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:1939118-Animals,
pubmed-meshheading:1939118-Binding Sites,
pubmed-meshheading:1939118-Calcium,
pubmed-meshheading:1939118-Carbohydrate Sequence,
pubmed-meshheading:1939118-Carrier Proteins,
pubmed-meshheading:1939118-Cations,
pubmed-meshheading:1939118-Chromatography, Gel,
pubmed-meshheading:1939118-Crystallography,
pubmed-meshheading:1939118-DNA Mutational Analysis,
pubmed-meshheading:1939118-Lanthanum,
pubmed-meshheading:1939118-Lectins,
pubmed-meshheading:1939118-Mannose-Binding Lectins,
pubmed-meshheading:1939118-Molecular Sequence Data,
pubmed-meshheading:1939118-Molecular Weight,
pubmed-meshheading:1939118-Peptide Fragments,
pubmed-meshheading:1939118-Peptide Mapping,
pubmed-meshheading:1939118-Rats,
pubmed-meshheading:1939118-Recombinant Proteins,
pubmed-meshheading:1939118-Structure-Activity Relationship
|
| pubmed:year |
1991
|
| pubmed:articleTitle |
Physical characterization and crystallization of the carbohydrate-recognition domain of a mannose-binding protein from rat.
|
| pubmed:affiliation |
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|