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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1999-2-5
|
| pubmed:abstractText |
Fibroblast growth factor-16 (FGF-16) is the most recent member of the FGF family to be cloned. Since the biologic activity of rat FGF-16 (rFGF-16) was unknown, and this protein has no apparent signal sequence, we transformed its entire cDNA into Escherichia coli for high-level expression and further characterization of this novel protein. An attempt was made to purify the expressed protein from the supernatant of mechanically lysed cells using sequential cation-exchange chromatography. This resulted in a gradual loss of the protein as precipitate throughout the purification process. In addition to precipitation during purification, sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the partially purified materials showed a cluster of protein bands around 20k to 29k. Sequence analysis of the major bands indicated that two N-terminal truncations had occurred, during E. coli fermentation, purification, or both. The largest truncation resulted in the removal of the 34 N-terminal amino acids, including the initiation codon methionine. We cloned d34 rFGF-16, expressed the gene in E. coli, and developed a purification process for this form. Even with this truncated form, precipitation was a problem. We were largely able to overcome this problem, however, by including EDTA throughout the purification process. We have characterized the structure of purified d34 rFGF-16 extensively using circular dichroism, Fourier transform infrared spectroscopy, and sedimentation velocity analysis. These studies revealed that the protein has a distinct tertiary structure, consists primarily of beta-strands, has a weak tendency to self-associate, and is fairly extended. We then performed biologic assays which showed that d34 rFGF-16 induces oligodendrocyte proliferation in vitro, and induces hepatocellular proliferation and increased liver weight in vivo. In summary, FGF-16, a novel FGF family member, has both unique structural and biological properties.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jan
|
| pubmed:issn |
0003-9861
|
| pubmed:author |
pubmed-author:ArakawaTT,
pubmed-author:BahriAA,
pubmed-author:ChiricaMM,
pubmed-author:DanilenkoD MDM,
pubmed-author:DelaneyJJ,
pubmed-author:HaraSS,
pubmed-author:HerndayNN,
pubmed-author:HillDD,
pubmed-author:ItohNN,
pubmed-author:KonishiMM,
pubmed-author:LOJJ,
pubmed-author:MartinFF,
pubmed-author:MontestruqueSS,
pubmed-author:OERIUSS,
pubmed-author:PhilpJ RJR,
pubmed-author:SpeakmanJJ,
pubmed-author:TalvenheimoJJ,
pubmed-author:ZhangMM
|
| pubmed:copyrightInfo |
Copyright 1999 Academic Press.
|
| pubmed:issnType |
Print
|
| pubmed:day |
1
|
| pubmed:volume |
361
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
34-46
|
| pubmed:dateRevised |
2004-11-18
|
| pubmed:meshHeading |
pubmed-meshheading:9882426-3T3 Cells,
pubmed-meshheading:9882426-Animals,
pubmed-meshheading:9882426-Biological Assay,
pubmed-meshheading:9882426-Circular Dichroism,
pubmed-meshheading:9882426-Crosses, Genetic,
pubmed-meshheading:9882426-Female,
pubmed-meshheading:9882426-Fibroblast Growth Factors,
pubmed-meshheading:9882426-Hydrolysis,
pubmed-meshheading:9882426-Injections, Intraperitoneal,
pubmed-meshheading:9882426-Keratinocytes,
pubmed-meshheading:9882426-Mice,
pubmed-meshheading:9882426-Mice, Inbred BALB C,
pubmed-meshheading:9882426-Mice, Inbred C57BL,
pubmed-meshheading:9882426-Mice, Inbred DBA,
pubmed-meshheading:9882426-Oligodendroglia,
pubmed-meshheading:9882426-Peptide Hydrolases,
pubmed-meshheading:9882426-Rats,
pubmed-meshheading:9882426-Recombinant Proteins,
pubmed-meshheading:9882426-Spectroscopy, Fourier Transform Infrared,
pubmed-meshheading:9882426-Structure-Activity Relationship,
pubmed-meshheading:9882426-Temperature
|
| pubmed:year |
1999
|
| pubmed:articleTitle |
Recombinant rat fibroblast growth factor-16: structure and biological activity.
|
| pubmed:affiliation |
Amgen Inc., Amgen Center, Thousand Oaks, California, 91320, USA. ddanilen@amgen.com
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| pubmed:publicationType |
Journal Article
|