The substrate specificity profile of human granzyme A.
The exact biological function of granzyme A, a granule-associated serine protease belonging to the tryptase family of proteases, is still a matter of debate because conflicting roles have been suggested, such as initiation of caspase-independent apoptosis-like cell death and endogenous modulation of inflammatory processes. In contrast to its well-studied family member, granzyme B, far less is known about the physiological targets of granzyme A. Using an N-terminal peptide-centric proteomics technology, the substrate specificity of human granzyme A was extensively characterized at the level of macromolecular protein substrates. Overall, more than 260 cleavage sites, almost exclusively favoring basic residues at the P1 position, in approximately 200 unique protein substrates, including the well-known in vitro substrates APEX-endonuclease 1 and different histones, were identified. Further substrate characterization was used to delineate physical properties in the substrate specificity profiles, which further highlights important aspects in protease/substrate biology.
Van Damme
Petra
P
Department of Medical Protein Research, Flanders Interuniversity Institute for Biotechnology, Ghent, Belgium. petra.vandamme@vib-ugent.be
Maurer-Stroh
Sebastian
S
Hao
Han
H
Colaert
Niklaas
N
Timmerman
Evy
E
Eisenhaber
Frank
F
Vandekerckhove
Joël
J
Gevaert
Kris
K
D000107:Acetylation; D000595:Amino Acid Sequence; D024361:Amino Acids, Basic / Q000737:chemistry / Q000378:metabolism; D030562:Databases, Protein; D053804:Granzymes / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry
D024361:Amino Acids, Basic; D010446:Peptide Fragments; D020543:Proteome; D011994:Recombinant Proteins; D053804:Granzymes; C509212:GZMA protein, human