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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1997-4-29
|
| pubmed:abstractText |
Plasminogen binding proteins have been described both for Gram positive and Gram negative bacteria. In the present work we describe the purification and characterization of a plasminogen binding protein from Haemophilus influenzae (strain HI-23459). Bacteria were sonicated in order to solubilize plasminogen-binding proteins. The supernatant was subjected to affinity chromatography on plasminogen kringle-4 fragment bound to Sepharose 4B and subsequently processed by ion-exchange chromatography on DEAE-Sepharose CL-6B. Characterization of the protein by SDS-PAGE displayed a single band with a molecular mass of about 55,000, both prior to and after reduction. The purified protein stimulates tPA (tissue plasminogen activator) catalysed plasminogen activation by a factor of approximately 300, mainly due to a decrease in K(m). Antibodies were raised in rabbits and used in quantitative and qualitative analysis. However, using a FITC-conjugate we failed to demonstrate the presence of the purified protein on the surface of intact bacteria. The corresponding gene was isolated from a lambda EMBL3 phage library prepared from chromosomal DNA from the same H. influenzae strain, using an oligonucleotide probe based on the NH2-terminal amino acid sequence. An open reading frame corresponding to 472 amino acid was found. The amino acid sequence of the translated gene demonstrates 97% identity with the recently published sequence from aspartate ammonia lyase (aspartase) from H. influenzae. Enzymatic analysis of the purified protein revealed a high aspartase activity.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Aspartate Ammonia-Lyase,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Plasminogen,
http://linkedlifedata.com/resource/pubmed/chemical/Tissue Plasminogen Activator,
http://linkedlifedata.com/resource/pubmed/chemical/plasminogen-binding protein...
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| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0006-3002
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
13
|
| pubmed:volume |
1324
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
182-90
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:9092705-Amino Acid Sequence,
pubmed-meshheading:9092705-Aspartate Ammonia-Lyase,
pubmed-meshheading:9092705-Bacterial Proteins,
pubmed-meshheading:9092705-Base Sequence,
pubmed-meshheading:9092705-Carrier Proteins,
pubmed-meshheading:9092705-Chromatography, Affinity,
pubmed-meshheading:9092705-Cloning, Molecular,
pubmed-meshheading:9092705-Genes, Bacterial,
pubmed-meshheading:9092705-Haemophilus influenzae,
pubmed-meshheading:9092705-Kinetics,
pubmed-meshheading:9092705-Molecular Sequence Data,
pubmed-meshheading:9092705-Molecular Weight,
pubmed-meshheading:9092705-Plasminogen,
pubmed-meshheading:9092705-Protein Binding,
pubmed-meshheading:9092705-Sequence Analysis, DNA,
pubmed-meshheading:9092705-Sequence Homology, Amino Acid,
pubmed-meshheading:9092705-Sequence Homology, Nucleic Acid,
pubmed-meshheading:9092705-Tissue Plasminogen Activator
|
| pubmed:year |
1997
|
| pubmed:articleTitle |
Purification and characterisation of a plasminogen-binding protein from Haemophilus influenzae. Sequence determination reveals identity with aspartase.
|
| pubmed:affiliation |
Department of Clinical Chemistry, Karolinska Hospital, Stockholm, Sweden.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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