1LY8

The crystal structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability and insight into modelling of protein structures

Summary for 1LY8

• Entry DOI: 10.2210/pdb1ly8/pdb
• Descriptor: Peroxidase, 2-acetamido-2-deoxy-beta-D-glucopyranose, beta-D-mannopyranose, ... (8 entities in total)
• Functional Keywords: peroxidase, mutant, thermostability, coprinus cinereus, oxidoreductase
• Biological source: Coprinopsis cinerea
• Cellular location: Secreted: P28314
• Total number of polymer chains: 2
• Total formula weight: 74616.33

Authors

Houborg, K.,Harris, P.,Poulsen, J.-C.N.,Svendsen, A.,Schneider, P.,Larsen, S. (deposition date: 2002-06-07, release date: 2002-06-14, Last modification date: 2024-10-09)

Primary citation

Houborg, K.,Harris, P.,Poulsen, J.C.,Schneider, P.,Svendsen, A.,Larsen, S.
The structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability.
Acta Crystallogr.,Sect.D, 59:997-1003, 2003

PubMed Abstract: Seven amino-acid substitutions introduced into the 343 amino-acid-long sequence of Coprinus cinereus peroxidase (CiP) led to a mutant enzyme (TS-rCiP) which is more stable than the native enzyme at higher temperature, pH and hydrogen peroxide concentrations. It is therefore more suitable for industrial applications. A structure determination was conducted on a deglycosylated but still active form of TS-rCiP based on X-ray diffraction data to 2.05 A resolution measured on a crystal cooled to 100 K and refined to R = 0.202 and R(free) = 0.249. The increased stability of the TS-rCiP enzyme can be understood from the structural changes of the TS-rCiP structure revealed by a comparative analysis with other known CiP structures. One of the more significant changes caused by three of the substitutions, I49S, V53A and T121A, is the conversion of a hydrophobic pocket into a hydrophilic pocket with associated changes in the water structure and the hydrogen-bonding interactions. The E239G substitution, which gives rise to increased thermostability at high pH, creates changes in the water structure and in the orientation of a phenylalanine (Phe236) in its vicinity. The three substitutions M166F, M242 and Y242F introduced to increase the oxidative stability do not introduce any structural changes.

PubMed: 12777761
DOI: 10.1107/S0907444903006784

Experimental method

X-RAY DIFFRACTION (2.05 Å)