2E1P

Crystal structure of pro-Tk-subtilisin

Summary for 2E1P

• Entry DOI: 10.2210/pdb2e1p/pdb
• Descriptor: Tk-subtilisin, CALCIUM ION (3 entities in total)
• Functional Keywords: subtilisin, serine protease, precursor, hydrolase
• Biological source: Thermococcus kodakarensis
• Cellular location: Secreted: P58502
• Total number of polymer chains: 1
• Total formula weight: 41632.79

Authors

Tanaka, S.,Saito, K.,Chon, H.,Matsumura, H.,Koga, Y.,Takano, K.,Kanaya, S. (deposition date: 2006-10-27, release date: 2007-01-16, Last modification date: 2024-10-23)

Primary citation

Tanaka, S.,Saito, K.,Chon, H.,Matsumura, H.,Koga, Y.,Takano, K.,Kanaya, S.
Crystal structure of unautoprocessed precursor of subtilisin from a hyperthermophilic archaeon: evidence for Ca2+-induced folding
J.Biol.Chem., 282:8246-8255, 2007

PubMed Abstract: The crystal structure of an active site mutant of pro-Tk-subtilisin (pro-S324A) from the hyperthermophilic archaeon Thermococcus kodakaraensis was determined at 2.3 A resolution. The overall structure of this protein is similar to those of bacterial subtilisin-propeptide complexes, except that the peptide bond linking the propeptide and mature domain contacts with the active site, and the mature domain contains six Ca2+ binding sites. The Ca-1 site is conserved in bacterial subtilisins but is formed prior to autoprocessing, unlike the corresponding sites of bacterial subtilisins. All other Ca2+-binding sites are unique in the pro-S324A structure and are located at the surface loops. Four of them apparently contribute to the stability of the central alphabetaalpha substructure of the mature domain. The CD spectra, 1-anilino-8-naphthalenesulfonic acid fluorescence spectra, and sensitivities to chymotryptic digestion of this protein indicate that the conformation of pro-S324A is changed from an unstable molten globule-like structure to a stable native one upon Ca2+ binding. Another active site mutant, pro-S324C, was shown to be autoprocessed to form a propeptide-mature domain complex in the presence of Ca2+. The CD spectra of this protein indicate that the structure of pro-S324C is changed upon Ca2+ binding like pro-S324A but is not seriously changed upon subsequent autoprocessing. These results suggest that the maturation process of Tk-subtilisin is different from that of bacterial subtilisins in terms of the requirement of Ca2+ for folding of the mature domain and completion of the folding process prior to autoprocessing.

PubMed: 17237225
DOI: 10.1074/jbc.M610137200

Experimental method

X-RAY DIFFRACTION (2.3 Å)