From Proteopedia
(Difference between revisions)
proteopedia linkproteopedia link
|
|
| Line 1: |
Line 1: |
| - | [[Image:2z57.jpg|left|200px]] | + | {{Seed}} |
| | + | [[Image:2z57.png|left|200px]] |
| | | | |
| | <!-- | | <!-- |
| Line 9: |
Line 10: |
| | {{STRUCTURE_2z57| PDB=2z57 | SCENE= }} | | {{STRUCTURE_2z57| PDB=2z57 | SCENE= }} |
| | | | |
| - | '''Crystal structure of G56E-propeptide:S324A-subtilisin complex'''
| + | ===Crystal structure of G56E-propeptide:S324A-subtilisin complex=== |
| | | | |
| | | | |
| - | ==Overview==
| + | <!-- |
| - | Tk-subtilisin [the mature domain of Pro-Tk-subtilisin in active form (Gly70-Gly398)] from the hyperthermophilic archaeon Thermococcus kodakaraensis is matured from Pro-Tk-subtilisin [a subtilisin homologue from T. kodakaraensis in pro form (Gly1-Gly398)] upon autoprocessing and degradation of propeptide. Pro-Tk-subtilisin is characterized by extremely slow maturation at mild temperatures, but this maturation rate is greatly increased by a single Gly56-->Ser mutation in the propeptide region. To analyze the role of Gly56, which assumes a left-handed conformation, Pro-Tk-subtilisin variants with complete amino acid substitutions at Gly56 were constructed. A comparison of their halo-forming activities suggests that all variants, except for Pro-G56W [Pro-G56X, Pro-Tk-subtilisin with Gly56-->X mutation (X = any amino acid)], mature faster than WT. Pro-G56W and Pro-G56E with the lowest and highest maturation rates, respectively, among 19 variants, as well as WT and Pro-G56S, were overproduced, purified, and characterized. SDS-PAGE analyses and Tk-subtilisin activity assay indicated that their maturation rates increased in the order WT < or = Pro-G56W < Pro-G56S < Pro-G56E. The propeptides of these variants were also overproduced, purified, and characterized. The stability and inhibitory potency of these propeptides decreased in the order Tk-propeptide [propeptide of Tk-subtilisin (Gly1-Leu69)] > or = G56W-propeptide > G56S-propeptide > G56E-propeptide, indicating that they are inversely correlated with the maturation rates of Pro7-Tk-subtilisin and its derivatives. The crystal structures of these propeptides determined in complex with S324A-subtilisin indicate that the conformation of the propeptide is altered by the mutation, such that nonglycine residues at position 56 assume a right-handed conformation and hydrophobic interactions at the core region decrease. These results indicate that Gly56 is required in stabilizing the propeptide fold. Stabilization of this fold leads to strong binding of Tk-propeptide to Tk-subtilisin, high resistance of Tk-propeptide to proteolytic degradation, and slow maturation of Pro-Tk-subtilisin.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_17988685}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 17988685 is the PubMed ID number. |
| | + | --> |
| | + | {{ABSTRACT_PUBMED_17988685}} |
| | | | |
| | ==About this Structure== | | ==About this Structure== |
| Line 36: |
Line 40: |
| | [[Category: Subtilisin]] | | [[Category: Subtilisin]] |
| | [[Category: Thermococcus kodakaraensis]] | | [[Category: Thermococcus kodakaraensis]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 19:57:34 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 15:50:09 2008'' |
Revision as of 12:50, 29 July 2008
Template:STRUCTURE 2z57
Crystal structure of G56E-propeptide:S324A-subtilisin complex
Template:ABSTRACT PUBMED 17988685
About this Structure
2Z57 is a Protein complex structure of sequences from Thermococcus kodakarensis. Full crystallographic information is available from OCA.
Reference
Requirement of left-handed glycine residue for high stability of the Tk-subtilisin propeptide as revealed by mutational and crystallographic analyses., Pulido MA, Tanaka S, Sringiew C, You DJ, Matsumura H, Koga Y, Takano K, Kanaya S, J Mol Biol. 2007 Dec 14;374(5):1359-73. Epub 2007 Oct 17. PMID:17988685
Page seeded by OCA on Tue Jul 29 15:50:09 2008
