4K1S

Gly-Ser-SplB protease from Staphylococcus aureus at 1.96 A resolution

Summary for 4K1S

• Entry DOI: 10.2210/pdb4k1s/pdb
• Related: 2AS9 2VID 2W7S 4K1T
• Descriptor: Serine protease SplB (2 entities in total)
• Functional Keywords: chymotrypsin-like fold, serine protease, extracellular, hydrolase
• Biological source: Staphylococcus aureus
• Cellular location: Secreted : Q2FXC3
• Total number of polymer chains: 2
• Total formula weight: 45092.43

Authors

Zdzalik, M.,Pustelny, K.,Stec-Niemczyk, J.,Cichon, P.,Czarna, A.,Popowicz, G.,Drag, M.,Wladyka, B.,Potempa, J.,Dubin, A.,Dubin, G. (deposition date: 2013-04-05, release date: 2014-04-16, Last modification date: 2023-11-08)

Primary citation

Pustelny, K.,Zdzalik, M.,Stach, N.,Stec-Niemczyk, J.,Cichon, P.,Czarna, A.,Popowicz, G.,Mak, P.,Drag, M.,Salvesen, G.S.,Wladyka, B.,Potempa, J.,Dubin, A.,Dubin, G.
Staphylococcal SplB Serine Protease Utilizes a Novel Molecular Mechanism of Activation.
J.Biol.Chem., 289:15544-15553, 2014

PubMed Abstract: Staphylococcal SplB protease belongs to the chymotrypsin family. Chymotrypsin zymogen is activated by proteolytic processing at the N terminus, resulting in significant structural rearrangement at the active site. Here, we demonstrate that the molecular mechanism of SplB protease activation differs significantly and we characterize the novel mechanism in detail. Using peptide and protein substrates we show that the native signal peptide, or any N-terminal extension, has an inhibitory effect on SplB. Only precise N-terminal processing releases the full proteolytic activity of the wild type analogously to chymotrypsin. However, comparison of the crystal structures of mature SplB and a zymogen mimic show no rearrangement at the active site whatsoever. Instead, only the formation of a unique hydrogen bond network, distant form the active site, by the new N-terminal glutamic acid of mature SplB is observed. The importance of this network and influence of particular hydrogen bond interactions at the N terminus on the catalytic process is demonstrated by evaluating the kinetics of a series of mutants. The results allow us to propose a consistent model where changes in the overall protein dynamics rather than structural rearrangement of the active site are involved in the activation process.

PubMed: 24713703
DOI: 10.1074/jbc.M113.507616

Experimental method

X-RAY DIFFRACTION (1.96 Å)