کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
34689 45039 2012 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Production of bioactive extracellular domain of pig and chicken activin type IIB receptors in Pichia pastoris
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Production of bioactive extracellular domain of pig and chicken activin type IIB receptors in Pichia pastoris
چکیده انگلیسی

Myostatin (MSTN) is a potent negative regulator for skeletal muscle growth, and binds to activin type IIB receptor (ActRIIB) for its cellular signal transduction. Administration of the extracellular domain of ActRIIB (ActRIIB-ECD) improved skeletal muscle growth in laboratory animals, suggesting that ActRIIB-ECD can be a useful pharmacological agent to improve skeletal muscle growth of meat-producing animals. In the current study, pig and chicken ActRIIB-ECDs were produced in the Pichia pastoris GS115, and the recombinant proteins were purified from induced culture media by Ni-NTA affinity chromatography. The digestion of pig and chicken ActRIIB-ECDs with PNGase F and glycoprotein staining demonstrated an N-linked glycosylation of these recombinant proteins. Glycoprotein staining also indicated an additional presence of glycosylation in chicken ActRIIB-ECD. Both the pig and chicken ActRIIB-ECDs were shown to inhibit MSTN activity in a reporter gene assay system in vitro. When MSTN-inhibitory potencies were compared by analyzing EC50 values, no difference in MSTN-inhibitory potency was observed between the glycosylated and N-deglycosylated forms of pig or chicken ActRIIB-ECD, suggesting that glycosylation does not affect the bioactivity of ActRIIB-ECD. MSTN-inhibitory potency of chicken ActRIIB-ECD was greater (P < 0.01) than that of pig ActRIIB-ECD. Results of this study demonstrate that bioactive pig and chicken ActRIIB-ECDs can be produced from P. pastoris. In addition, the study indicates that the N-glycosylation status of ActRIIB-ECD does not affect its bioactivity in vitro.


► Extracellular domains (ECD) of chicken and pig ActRIIB are expressed in Pichia pastoris.
► Chicken and pig ActRIIB-ECD are N-glycosylated.
► Chicken ActRIIB appears to have O-glycosylation.
► N-glycosylation has no effect on the binding of ActRIIB-ECD to myostatin.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Process Biochemistry - Volume 47, Issue 1, January 2012, Pages 139–146
نویسندگان
, , ,