| Purification of recombinant growth hormone by clear native gels for conformational analyses: preservation of conformation and receptor binding. | |
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MedLine Citation:
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PMID: 20238132 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Most protein preparations require purification steps prior to biophysical analysis assessing protein stability, secondary structure and degree of folding. It was, therefore, the aim of this study to develop a system to separate and purify a protein from a commercially available medicinal product, recombinant human growth hormone (rhGH) and show preservation of conformation and function following the gel-based procedure. The rhGH was run on clear native (CN) gels and recovered from the gels by electroelution using D-Tube Dialyzer Midi under rigorous cooling. Melting point studies indicated preservation of the structural integrity. This finding was confirmed by synchrotron radiation circular dichroism spectroscopy (SRCD) revealing an identical folding pattern for the sample before and after electrophoretic separation and purification. Synchrotron small-angle X-ray scattering (SAXS) indicated that the sample was folded and monomeric, both before and after separation and purification, and that its shape corresponded well to the known crystal structure of GH. Binding properties of rhGH to a receptor-model system before and after clear native electrophoresis were comparable. This analytical and preparative approach to purify and concentrate a protein preserving conformation and function may be helpful for many applications in analytical, protein and stereochemistry. |
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Authors:
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Wei-Qiang Chen; Anita Salmazo; Matti Myllykoski; Björn Sjöblom; Martin Bidlingmaier; Arnold Pollak; Peter Baumgärtel; Kristina Djinovic-Carugo; Petri Kursula; Gert Lubec |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-03-19 |
Journal Detail:
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Title: Amino acids Volume: 39 ISSN: 1438-2199 ISO Abbreviation: Amino Acids Publication Date: 2010 Aug |
Date Detail:
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Created Date: 2010-07-23 Completed Date: 2010-10-28 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9200312 Medline TA: Amino Acids Country: Austria |
Other Details:
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Languages: eng Pagination: 859-69 Citation Subset: IM |
Affiliation:
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Department of Pediatrics, Medical University of Vienna, Vienna, Austria. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Electrophoresis, Polyacrylamide Gel Growth Hormone / chemistry*, genetics, isolation & purification*, metabolism Humans Molecular Sequence Data Peptide Mapping Protein Binding Protein Conformation Protein Folding Receptors, Somatotropin / chemistry*, metabolism Recombinant Proteins / chemistry, genetics, isolation & purification, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Receptors, Somatotropin; 0/Recombinant Proteins; 9002-72-6/Growth Hormone |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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