Literature summary for 3.4.21.79 extracted from

Ho, P.; Ede, C.; Chen, Y.Y.
Modularly constructed synthetic granzyme B molecule enables interrogation of intracellular proteases for targeted cytotoxicity (2017), ACS Synth. Biol., 6, 1484-1495 .

Search for more literature for 3.4.21.79

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of a protein-based therapeutic platform, termed cytoplasmic oncoprotein verifier and response trigger (COVERT), which enables the interrogation of intracellular proteases to trigger targeted cytotoxicity. COVERT molecules consist of the cytotoxic protein granzyme B (GrB) fused to an inhibitory N-terminal peptide, which can be removed by researcher-specified proteases to activate GrB function. Fusion of a small ubiquitin-like modifier 1 (SUMO1) protein to GrB yields a SUMO-GrB molecule that is specifically activated by the cancer-associated sentrin-specific protease 1 (SENP1). SUMO-GrB selectively triggers apoptotic phenotypes in HEK293T cells that overexpress SENP1, and it is highly sensitive to different SENP1 levels across cell lines. The rational design of additional COVERT molecules responsive to enterokinase (EK) and tobacco etch virus protease (TEVp) highlight the COVERT platform's modularity and adaptability to diverse protease targets. Primary human T cells (Jurkat cells) can express, package, traffic, and deliver engineered GrB molecules in response to antigen stimulation. The COVERT platform, ENLYFQ-GrB is evaluated for its ability to selectively mediate cytotoxicity against HEK293T cells expressing TEVp. In contrast to SENP1, TEVp is not mammalian in origin, and its forced expression mimics a state of viral infection. Transient expression of GrB in HEK293T cells resulted in marked changes in cell physiology regardless of whether the cells co-expressed TEVp. Method evaluation supporting the modularity and versatility of the COVERT platform for targeting proteolytic markers of a variety of disease states, overview	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P10144	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	endogenous GrB is produced as a zymogen bearing an N-terminal Gly-Glu dipeptide that prevents the formation of a functional catalytic triad. Upon packaging into lytic granules inside the immune cell, GrB is processed by the dipeptidyl peptidase cathepsin C (CatC), which cleaves off GrB's Gly-Glu dipeptide and frees the newly N-terminal Ile16 residue to insert into the interior of the molecule and form a salt bridge with Asp194. The resulting conformational change enables the simultaneous generation of an oxyanion hole and maturation of the active-site S1 pocket. Endogenous GrB is activated prior to its release from the lytic granules of T cells and NK cells	Homo sapiens

Synonyms

Synonyms	Comment	Organism
GrB	-	Homo sapiens

General Information

General Information	Comment	Organism
physiological function	granzyme B (GrB) is the initiator of multiple pro-apoptotic pathways and serves as the principle cytotoxic molecule deployed by T cells and natural killer (NK) cells to eliminate target cells. Endogenous GrB is produced as a zymogen bearing an N-terminal Gly-Glu dipeptide that prevents the formation of a functional catalytic triad. Upon packaging into lytic granules inside the immune cell, GrB is processed by the dipeptidyl peptidase cathepsin C (CatC), which cleaves off GrB's Gly-Glu dipeptide and frees the newly N-terminal Ile16 residue to insert into the interior of the molecule and form a salt bridge with Asp194. The resulting conformational change enables the simultaneous generation of an oxyanion hole and maturation of the active-site S1 pocket. Since endogenous GrB is activated prior to its release from the lytic granules of T cells and NK cells, it indiscriminately kills any target cell it enters and does not independently ascertain the identity of the target cell. Target-cell identification is established exclusively at the cell surface via receptor-antigen interactions	Homo sapiens

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)