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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1992-1-21
|
| pubmed:abstractText |
The lactose-assimilating yeast, Kluyveromyces lactis, has been developed as a microbial host for the synthesis and secretion of human proteins. Here, we report the use of multi-copy vectors based on the 2 mu-like plasmid pKD1 from Kluyveromyces drosophilarum [Chen et al., Nucleic Acids Res. 14 (1986) 4471-4481] for the secretion of recombinant human interleukin-1 beta (reIL-1 beta). High levels of reIL-1 beta were secreted into the growth medium when the structural gene was fused in-frame to a synthetic secretion signal derived from the 'pre'-region of the K. lactis killer toxin. N-terminal sequencing of the excreted protein showed highly efficient (greater than 95%) maturation of the signal sequence. Synthesis as prepro-IL-1 beta, the 'pro'-sequence being derived from the human serum albumin-encoding gene, resulted in equally efficient secretion of mature IL-1 beta. Cytoplasmic production of Met-IL-1 beta, without a secretion signal, was found to be toxic to K. lactis. As in Saccharomyces cerevisiae [Baldari et al., EMBO J. 6 (1987) 229-234], but unlike native human IL-1 beta, K. lactis reIL-1 beta is glycosylated. This glycosylation led to a 95% loss of its biological activity. Removal of the carbohydrate chains by endo-beta-N-acetyl-glucosamidase H treatment fully restored the biological activity. A modified form of IL-1 beta (Asn7----Gln7), in which the unique site for Asn-linked glycosylation was deleted, exhibited the same biological activity as native IL-1 beta. The level of secretion of mature recombinant IL-1 beta ws glycosylation-independent.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Interleukin-1,
http://linkedlifedata.com/resource/pubmed/chemical/Killer Factors, Yeast,
http://linkedlifedata.com/resource/pubmed/chemical/Mycotoxins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/zymocin
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0378-1119
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
15
|
| pubmed:volume |
107
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
N
|
| pubmed:pagination |
285-95
|
| pubmed:dateRevised |
2009-11-19
|
| pubmed:meshHeading |
pubmed-meshheading:1748298-Amino Acid Sequence,
pubmed-meshheading:1748298-Base Sequence,
pubmed-meshheading:1748298-Blotting, Northern,
pubmed-meshheading:1748298-Blotting, Southern,
pubmed-meshheading:1748298-Blotting, Western,
pubmed-meshheading:1748298-Gene Expression,
pubmed-meshheading:1748298-Genetic Vectors,
pubmed-meshheading:1748298-Glycosylation,
pubmed-meshheading:1748298-Humans,
pubmed-meshheading:1748298-Interleukin-1,
pubmed-meshheading:1748298-Killer Factors, Yeast,
pubmed-meshheading:1748298-Kinetics,
pubmed-meshheading:1748298-Kluyveromyces,
pubmed-meshheading:1748298-Molecular Sequence Data,
pubmed-meshheading:1748298-Mycotoxins,
pubmed-meshheading:1748298-Plasmids,
pubmed-meshheading:1748298-Protein Processing, Post-Translational,
pubmed-meshheading:1748298-Recombinant Fusion Proteins
|
| pubmed:year |
1991
|
| pubmed:articleTitle |
High-level secretion of correctly processed recombinant human interleukin-1 beta in Kluyveromyces lactis.
|
| pubmed:affiliation |
Rhône Poulenc Rorer, Biotechnology, Department, Vitry, France.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|