Receptor-binding domain of human α2-macroglobulin Expression, folding and biochemical characterization of a high-affinity recombinant derivative
| Autor: | Kåre Lehmann Nielsen, Lars Sottrup-Jensen, Hans Christian Thøgersen, Thor Las Holtet, Michael Etzerodt, Søren K. Moestrup, Jørgen Gliemann |
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| Rok vydání: | 1994 |
| Předmět: |
Protein Folding
Placenta Molecular Sequence Data Biophysics Gene Expression Biology Cleavage (embryo) Binding Competitive Biochemistry α2-Macroglobulin receptor law.invention Structural Biology law Escherichia coli Genetics Humans alpha-Macroglobulins Amino Acid Sequence Amino Acids Receptors Immunologic Binding site Receptor Molecular Biology Peptide sequence Polyacrylamide gel electrophoresis Binding Sites Base Sequence Gene Transfer Techniques Cell Biology Peptide Fragments Recombinant Proteins Macroglobulin α-Macroglobulin Recombinant DNA Protein expression Domain structure Electrophoresis Polyacrylamide Gel Female Protein folding Low Density Lipoprotein Receptor-Related Protein-1 Plasmids |
| Zdroj: | FEBS Letters. 344:242-246 |
| ISSN: | 0014-5793 |
| Popis: | A recombinant version of the receptor binding domain (RBDv) of human alpha 2-macroglobulin (alpha 2M) has been expressed in E. coli and refolded using a novel iterative procedure. RBDv (Val1299-Ala1451) is extended by 15 residues at the N-terminal side of the Lys1313-Glu papain cleavage site in human alpha 2M. RBDv contains the intra-chain bridge Cys1329-Cys1444 and is soluble and monomeric. Competition experiments with 125I-labelled methylamine-treated alpha 2M reveal that RBDv binds to the placental receptor for transformed alpha 2M with a Kd of 8 nM, i.e. the binding affinity of RBDv is of the same order of magnitude as the intrinsic affinity for binding of one domain in transformed alpha 2M to one receptor molecule. |
| Databáze: | OpenAIRE |
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