Abstract
The cell death–inducing serine protease granzyme A (GzmA) has a unique disulfide-linked quaternary structure. The structure of human GzmA bound to a tripeptide CMK inhibitor, determined at a resolution of 2.4 Å, reveals that the oligomeric state contributes to substrate selection by limiting access to the active site for potential macromolecular substrates and inhibitors. Unlike other serine proteases, tetrapeptide substrate preferences do not correlate well with natural substrate cleavage sequences. This suggests that the context of the cleavage sequence within a macromolecular substrate imposes another level of selection not observed with the peptide substrates. Modeling of inhibitors bound to the GzmA active site shows that the dimer also contributes to substrate specificity in a unique manner by extending the active-site cleft. The crystal structure, along with substrate library profiling and mutagenesis, has allowed us to identify and rationally manipulate key components involved in GzmA substrate specificity.
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Acknowledgements
The authors would like to express thanks to J. Holton for technical assistance during data collection at the ALS beam line 8.3.1. We would also like to acknowledge the Macromolecular Structure Group at the University of California, San Francisco for computational support. This work was supported in part by a US National Institutes of Health post-doctoral training grant to J.K.B. and grants from the US National Institutes of Health to C.S.C. and R.J.F.
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Bell, J., Goetz, D., Mahrus, S. et al. The oligomeric structure of human granzyme A is a determinant of its extended substrate specificity. Nat Struct Mol Biol 10, 527–534 (2003). https://doi.org/10.1038/nsb944
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DOI: https://doi.org/10.1038/nsb944
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