4HD7

Crystal Structure of Tyrosinase from Bacillus megaterium V218G mutant soaked in CuSO4

Summary for 4HD7

• Entry DOI: 10.2210/pdb4hd7/pdb
• Related: 4HD4 4HD6 4HDA
• Descriptor: Tyrosinase, COPPER (II) ION (3 entities in total)
• Functional Keywords: tyrosinase, type 3 copper protein, oxidoreductase
• Biological source: Bacillus megaterium
• Total number of polymer chains: 2
• Total formula weight: 70678.97

Authors

Goldfeder, M.,Kanteev, M.,Adir, N.,Fishman, A. (deposition date: 2012-10-02, release date: 2013-01-23, Last modification date: 2024-02-28)

Primary citation

Goldfeder, M.,Kanteev, M.,Adir, N.,Fishman, A.
Influencing the monophenolase/diphenolase activity ratio in tyrosinase.
Biochim.Biophys.Acta, 1834:629-633, 2013

PubMed Abstract: Tyrosinase is a type 3 copper enzyme with great potential for production of commercially valuable diphenols from monophenols. However, the use of tyrosinase is limited by its further oxidation of diphenols to quinones. We recently determined the structure of the Bacillus megaterium tyrosinase revealing a residue, V218, which we proposed to take part in positioning of substrates within the active site. In the structure of catechol oxidase from Ipomoea batatas, the lack of monophenolase activity was attributed to the presence of F261 near CuA. Consequently, we engineered two variants, V218F and V218G. V218F was expected to have a decreased monophenolase activity, due to the bulky residue extending into the active site. Surprisingly, both V218F and V218G exhibited a 9- and 4.4-fold higher monophenolase/diphenolase activity ratio, respectively. X-ray structures of variant V218F display a flexibility of the phenylalanine residue along with an adjacent histidine, which we propose to be the source of the change in activity ratio.

PubMed: 23305929
DOI: 10.1016/j.bbapap.2012.12.021

Experimental method

X-RAY DIFFRACTION (2.1 Å)