3G01

Structure of GrC mutant E192R/E193G

Summary for 3G01

• Entry DOI: 10.2210/pdb3g01/pdb
• Related: 3FZZ
• Descriptor: Granzyme C (2 entities in total)
• Functional Keywords: hydrolase, cytolysis, protease, serine protease, zymogen
• Biological source: Mus musculus (mouse)
• Cellular location: Cytoplasmic granule: P08882
• Total number of polymer chains: 2
• Total formula weight: 50296.11

Authors

Buckle, A.M.,Kaiserman, D.,Whisstock, J.C. (deposition date: 2009-01-27, release date: 2009-03-17, Last modification date: 2024-10-30)

Primary citation

Kaiserman, D.,Buckle, A.M.,Van Damme, P.,Irving, J.A.,Law, R.H.P.,Matthews, A.Y.,Bashtannyk-Puhalovich, T.,Langendorf, C.,Thompson, P.,Vandekerckhove, J.,Gevaert, K.,Whisstock, J.C.,Bird, P.I.
Structure of granzyme C reveals an unusual mechanism of protease autoinhibition
Proc.Natl.Acad.Sci.USA, 106:5587-5592, 2009

PubMed Abstract: Proteases act in important homeostatic pathways and are tightly regulated. Here, we report an unusual structural mechanism of regulation observed by the 2.5-A X-ray crystal structure of the serine protease, granzyme C. Although the active-site triad residues adopt canonical conformations, the oxyanion hole is improperly formed, and access to the primary specificity (S1) pocket is blocked through a reversible rearrangement involving Phe-191. Specifically, a register shift in the 190-strand preceding the active-site serine leads to Phe-191 filling the S1 pocket. Mutation of a unique Glu-Glu motif at positions 192-193 unlocks the enzyme, which displays chymase activity, and proteomic analysis confirms that activity of the wild-type protease can be released through interactions with an appropriate substrate. The 2.5-A structure of the unlocked enzyme reveals unprecedented flexibility in the 190-strand preceding the active-site serine that results in Phe-191 vacating the S1 pocket. Overall, these observations describe a broadly applicable mechanism of protease regulation that cannot be predicted by template-based modeling or bioinformatic approaches alone.

PubMed: 19299505
DOI: 10.1073/pnas.0811968106

Experimental method

X-RAY DIFFRACTION (2.5 Å)