Expression and purification of recombinant human EGFL7 protein.
The secreted epidermal growth factor-like protein 7 (EGFL7) plays an important role in angiogenesis, especially in the recruitment of endothelial and smooth muscle cells to the site of the nascent vessel and their ordered assembly into functional vasculature. However, progress in the understanding of the underlying mechanisms is to date greatly hindered by the lack of recombinant EGFL7 protein in a stable, soluble, native state, thus preventing e.g. the characterization of the proposed functional receptor as well as investigation of additional biological effects of EGFL7. So far all attempts to produce sufficient amounts of recombinant EGFL7 protein by various groups have failed. In this study we describe a procedure for the expression and purification of human EGFL7 from Sf9 cells and for the first time provide means to isolate biologically functional EGFL7 protein in sufficient quantities for its further biological characterization. We believe that the availability of EGFL7 will greatly accelerate our understanding of the precise role of EGFL7 and the underlying molecular mechanisms of EGFL7 action in the fundamentally important process of angiogenesis.
Picuric
Srdjan
S
Institute of Biochemistry II, Johann Wolfgang Goethe University School of Medicine, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany.
Friedrich
Matthias
M
Oess
Stefanie
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002135:Calcium-Binding Proteins; D002460:Cell Line; D002465:Cell Movement; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002942:Circular Dichroism; D066255:EGF Family of Proteins; D016228:Endothelial Growth Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera / Q000378:metabolism
D002135:Calcium-Binding Proteins; D066255:EGF Family of Proteins; C527877:EGFL7 protein, human; D016228:Endothelial Growth Factors; D011994:Recombinant Proteins