Receptor‐binding domain of human α2‐macroglobulin Expression, folding and biochemical characterization of a high‐affinity recombinant derivative
- 16 May 1994
- journal article
- Published by Wiley in FEBS Letters
- Vol. 344 (2-3) , 242-246
- https://doi.org/10.1016/0014-5793(94)00349-1
Abstract
A recombinant version of the receptor binding domain (RBDv) of human α2‐macroglobulin (α2M) 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 α2M. RBDv contains the intra‐chain bridge Cys1329‐Cys1444 and is soluble and monomeric. Competition experiments with 125I‐labelled methylamine‐treated α2M reveal that RBDv binds to the placental receptor for transformed α2M with a K d 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 α2M to one receptor molecule.Keywords
This publication has 26 references indexed in Scilit:
- Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genesPublished by Elsevier ,2004
- Expression of a functional α‐macroglobulin receptor binding domain in Escherichia coliFEBS Letters, 1992
- Sequence‐specific binding of the N‐terminal three‐finger fragment of Xenopus transcription factor IIIA to the internal control region of a 5S RNA geneFEBS Letters, 1991
- Evidence that the newly cloned low‐density‐lipoprotein receptor related protein (LRP) is the α2‐macroglobulin receptorFEBS Letters, 1990
- Amino acid analysis: Determination of cysteine plus half-cystine in proteins after hydrochloric acid hydrolysis with a disulfide compound as additiveAnalytical Biochemistry, 1989
- A conserved region in .alpha.-macroglobulins participates in binding to the mammalian .alpha.-macroglobulin receptorBiochemistry, 1989
- Genetic Approach to Facilitate Purification of Recombinant Proteins with a Novel Metal Chelate AdsorbentNature Biotechnology, 1988
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Domain structure of human α2‐macroglobulinFEBS Letters, 1986
- Cell association and degradation of α2-macroglobulin-trypsin complexes in hepatocytes and adipocytesBiochimica et Biophysica Acta (BBA) - General Subjects, 1983