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Updated: Aug 23, 2026
A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
Published on: November 28, 2014
[Construction, expression and cell growth inhibition of translocating peptide/granzyme B fusion protein gene]
Jing Zhao1, Zhi Wang, Yan-ming Xu
1Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032 China.
Aim:
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.
Conclusion:
Expression of PE II-GrBa fusion protein can inhibit cell growth.
