[Construction, expression and cell growth inhibition of translocating peptide/granzyme B fusion protein gene].
OBJECTIVE
To investigate the inhibitory effect of translocating peptide/granzyme B fusion protein on cell growth.


METHODS
The translocating peptide gene of Pseudomonas exotoxin A (PE) was fused with active granzyme B gene by recombinant PCR to construct PE II-GrBa fusion protein gene. PE II-mGrBa with a mutation of serine to cystein at active center of GrB was used as negative control. The resulting PE II-GrBa and PE II-mGrBa genes were transiently transfected into mammalian cells via lipofectamine mediation. The effects of expression of PE II-GrBa gene on morphology and growth of transfected cells were detected by MTT colorimetry, TUNEL assay and indirect immunofluorescence staining.


RESULTS
Transient expression of PE II-GrBa resulted in cytoskeleton abnormality, cell growth inhibition, and apoptosis in some cells.


CONCLUSIONS
Expression of PE II-GrBa fusion protein can inhibit cell growth.
Zhao
Jing
J
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032 China.
Wang
Zhi
Z
Xu
Yan-ming
YM
Gui
Jun-hao
JH
Wen
Wei-hong
WH
Wang
Cheng-ji
CJ
Yang
An-gang
AG
D017209:Apoptosis; D049109:Cell Proliferation; D005098:Exotoxins / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D053804:Granzymes; D006367:HeLa Cells; D006801:Humans; D010446:Peptide Fragments / Q000235:genetics; D011549:Pseudomonas; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D005098:Exotoxins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases