TY  - JOUR
U1  - Zeitschriftenartikel, wissenschaftlich - begutachtet (reviewed)
A1  - Mellert, Kevin
A1  - Lechner, Stefan
A1  - Lüdeke, Manuel
A1  - Lamla, Markus
A1  - Möller, Peter
A1  - Kemkemer, Ralf
A1  - Scheffzek, Klaus
A1  - Kaufmann, Dieter
T1  - Restoring functional neurofibromin by protein transduction
JF  - Scientific reports
N2  - In Neurofibromatosis 1 (NF1) germ line loss of function mutations result in reduction of cellular neurofibromin content (NF1+/−, NF1 haploinsufficiency). The Ras-GAP neurofibromin is a very large cytoplasmic protein (2818 AA, 319 kDa) involved in the RAS-MAPK pathway. Aside from regulation of proliferation, it is involved in mechanosensoric of cells. We investigated neurofibromin replacement in cultured human fibroblasts showing reduced amount of neurofibromin. Full length neurofibromin was produced recombinantly in insect cells and purified. Protein transduction into cultured fibroblasts was performed employing cell penetrating peptides along with photochemical internalization. This combination of transduction strategies ensures the intracellular uptake and the translocation to the cytoplasm of neurofibromin. The transduced neurofibromin is functional, indicated by functional rescue of reduced mechanosensoric blindness and reduced RasGAP activity in cultured fibroblasts of NF1 patients or normal fibroblasts treated by NF1 siRNA. Our study shows that recombinant neurofibromin is able to revert cellular effects of NF1 haploinsuffiency in vitro, indicating a use of protein transduction into cells as a potential treatment strategy for the monogenic disease NF1.
Y1  - 2018
UN  - https://nbn-resolving.org/urn:nbn:de:bsz:rt2-opus4-21493
SN  - 2045-2322
SS  - 2045-2322
U6  - https://doi.org/10.1038/s41598-018-24310-5
DO  - https://doi.org/10.1038/s41598-018-24310-5
VL  - 8
IS  - Aufsatz 6171
SP  - 1
EP  - 9
S1  - 9
PB  - Macmillan Publishers Limited
CY  - London
ER  -