title: Expression and purification of biologically active rat bone morphogenetic protein-4 produced as inclusion bodies in recombinant Escherichia coli.
abstract: Rat bone morphogenetic protein-4 (rBMP-4) cDNA was cloned from rat osteoblasts by RT-PCR and expressed in E. coli. Monomeric, dimeric and polymeric forms of recombinant rat BMP-4 (rrBMP-4) were obtained from inclusion bodies after solubilization with urea. The dimer was separated from the remaining polymer and host cell contaminants using size exclusion chromatography. Furthermore, purified rrBMP-4 was stabilized at low urea concentration (40 mM) and at pH 8.5 through the addition of bovine serum albumin. Both, rrBMP-4 dimer and polymer were biologically active as tested by the induction of alkaline phosphatase activity in MC3T3-E1 cells.
authors.affiliation: Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Donaueschingenstrasse 13, 1200, Vienna, Austria. burkhard.kloesch@lbitrauma.org
authors.forename: Burkhard
authors.initials: B
authors.lastname: Klösch
authors.forename: Walter
authors.initials: W
authors.lastname: Fürst
authors.forename: Rudolf
authors.initials: R
authors.lastname: Kneidinger
authors.forename: Monika
authors.initials: M
authors.lastname: Schuller
authors.forename: Barbara
authors.initials: B
authors.lastname: Rupp
authors.forename: Asmita
authors.initials: A
authors.lastname: Banerjee
authors.forename: Heinz
authors.initials: H
authors.lastname: Redl
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D002479:Inclusion Bodies / Q000235:genetics; D010006:Osteoblasts / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: C525811:Bmp4 protein, rat; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D011994:Recombinant Proteins