Granzyme B signaling
Granzyme B is abundant serine protease, which is
responsible for rapid induction of caspase-dependent apoptosis, promoting caspase
activation directly and indirectly, through proteolysis of the Bcl-2 family proteins
[1], [2].
CD8+ T cells cytotoxicity is activated after cell infection, cancer transformation,
and allogenic antigens presentation by MHC class I. NK cells are activated after
recognition of allogenic or modified antigens on MHC class I-negative target cell surface
[3], [4], [5]. This results in NK or CD8+ T cells
polarization and maturation of cytotoxic granule inclusive Granzyme
B and Perforin. Synaptotagmin
VII [6], [7],
Rab-27A and Unc-13 homolog D
(Munc13-4) are essential regulators of cytotoxic granule
maturation and fusion with the plasma membrane [8], [9], [10]. These granules are localized in contact with the target cells and contain also
lysosome-resident proteins Lysosomal-associated membrane protein 1 and 2
(LAMP1 and LAMP2) and
CD63 [11]. Perforin
and Granzyme B release from cytotoxic granules into the
immunological synapse formed between effector cells [12].
Perforin allows Granzyme B
access to cytosolic and nuclear substrates [13], [14], [15]. In the cytoplasm human Granzyme B cleaves BH3
interacting domain death agonist (Bid) more efficiently than
it cleaves caspases [1]. Proteolysis of Bid by
Granzyme B results in the translocation of the C terminus of
Bid (tBid) to mitochondria. In mitochondria outer membrane
Granzyme B also cleaves anti-apoptotic Bcl-2 family protein
Myeloid cell leukemia sequence 1 (Mcl-1) [16], [17]. Those interactions induce the release of mitochondrial Cytochrome c
somatic (Cytochrome C) and Diablo homolog
(Smac/Diablo) into the cytosol [2].
Cytochrome C is involved in the apoptosome pathway together
with Apoptotic peptidase activating factor 1 (Apaf-1) and
activated Caspase 9 [18], [19].
Release of mitochondrial Smac/Diablo represses the
inhibitors of apoptosis in target cell, making caspases more readily activated directly
by Granzyme B [19].
On the other hand, Granzyme B directly processes
Caspase-8, and Caspase-10, and
then activates Caspase-3, and
Caspase-7. Caspase-3 is the
central effector caspase within the Granzyme B-initiated caspase cascade. It completes
maturation of Caspase-8 and
Caspase-10 and activates
Caspase-2, Caspase-6 and
Caspase-9 [1].
Granzyme B directly cleaves caspase substrates, like
nuclear Lamin B1 [20], Nuclear mitotic apparatus
protein 1 (NUMA1) [2], [21], Poly
(ADP-ribose) polymerase 1 (PARP-1) [22], [23], and cytoplasmic DNA fragmentation factor 45kDa alpha polypeptide
(ICAD) [24], and Rho-associated coiled-coil
containing protein kinase 2 (ROCK2) [25].
Caspases also cleave Lamin A/C [15].
Cleavage of structural components of nucleoskeleton,
Lamins and NUMA1, results in
nuclear integrity disruption [20], [26]. Degradation of
ICAD induces release of DNA fragmentation factor 40kDa beta
polypeptide (DFF40 (CAD)) that
degrades chromosomal DNA [24], [27]. Cleavage of
PARP-1 promotes apoptosis by preventing DNA repair-induced
survival [28]. Degradation of ROCK2 results in
membrane blebbing [2], [25]. Granzyme
B cleaves Tubulin alpha and induces
microtubule polymerization and cytoskeletal collapse [29]. Proteolysis of
Fibroblast growth factor receptor 1 (FGFR1) and
transmembrane Notch homolog 1 translocation-associated (NOTCH1
(NEXT)) results in target cell isolation from survival signals from
extracellular environment and enhances programmed cell death, apoptosis [22].
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