Lack of prodomain abolishes BDNF constitutive secretion by HEK 293 cells

• Autorzy: Ziemlinska E. , Skup M. , Czarkowska-Bauch J. , Bahr M. , Kugler S.
• Języki publikacji: EN

Abstrakty

EN

Brain-derived neurotrophic factor (BDNF) and its proBDNF precursor are released both in constitutive and activity-dependent manner. Prodomain itself proved to be necessary for BDNF targeting to regulated secretory pathway [Egan et al. (2003) Cell, Chen et al. (2005) J Neurosci] but its role in constitutive secretion is elusive. As mature BDNF (mBDNF) conveys trophic and prosurvival signals whereas proBDNF may convey growth inhibiting and death signals, an important issue arises: can we control the type of signal being triggered by BDNF? To verify this we cut off the prodomain and generated plasmid coding only for rat mBDNF. To test the constitutive mBDNF construct secretion we have chosen HEK 293 cell line. Two other plasmids coding either for proBDNF (template for both BDNF forms) or proBDNF protected from prodomain cleavage (source of proBDNF only), served as controls. BDNF secretion was evaluated with WB technique using antibodies detecting (1) both BDNF forms, (2) HA tag (mBDNF construct) and (3) MYC tag (proBDNF constructs). We found all three constructs being stably expressed in HEK cells. However, in contrast to both proBDNF constructs, which were secreted and detected in media fraction, mBDNF construct was revealed only in the cell lysate fraction, not being released to the media. This is the fi rst observation showing that mBDNF can be constitutively released only when accompanied by prodomain. Support: ASTF 211-00-2007 for EZ, Polish-German grant to MS and SK.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2009
• Tom: 69
• Numer: 3
• Opis fizyczny: p.307

Twórcy

autor

Ziemlinska E.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Skup M.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Czarkowska-Bauch J.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Bahr M.

• Department of Neurology, University Hospital Gottingen, Gottingen, Germany

autor

Kugler S.

• Department of Neurology, University Hospital Gottingen, Gottingen, Germany