Large scale protein production of the extracellular domain of the transforming growth factor-beta type II receptor using the Pichia pastoris expression system.
To study the (patho)physiological role of transforming growth factor-beta (TGF-beta), potent and selective inhibitors are necessary. Since TGF-beta signaling is initiated by the high affinity binding to the type II receptor (RII), the extracellular part of RII (solRII) can function as a TGF-beta antagonist. SolRII was cloned and large-scale protein synthesis was performed in the yeast Pichia pastoris expression system. Our results indicate that via this system, high levels of pure concentrated solRII can be obtained. Moreover, purified solRII is an active protein as shown by ELISA and bioassay. In conclusion, our large-scale protein expression procedure results in high quantities of purified solRII, which is a powerful tool to study the natural role of TGF-beta.
Scharstuhl
Alwin
A
Rheumatology Research Laboratory, Department of Rheumatology, University Medical Center Nijmegen, Geert Grooteplein zuid 26-28, 6525 GA, Nijmegen, The Netherlands. a.scharstuhl@ncmls.kun.nl
Glansbeek
Harrie
H
Vitters
Elly L
EL
van der Kraan
Peter M
PM
van den Berg
Wim B
WB
D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D007525:Isoelectric Focusing; D010843:Pichia / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053781:Transforming Growth Factor beta2
D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II