5GX9

PYP mutant - E46Q

Summary for 5GX9

• Entry DOI: 10.2210/pdb5gx9/pdb
• Descriptor: Photoactive yellow protein, 4'-HYDROXYCINNAMIC ACID (3 entities in total)
• Functional Keywords: pas, lov, photoreceptor, light sensor, lbhb, shb, chromophore, photoreceptor protein, receptor, sensory transduction, signaling protein
• Biological source: Halorhodospira halophila
• Total number of polymer chains: 1
• Total formula weight: 14051.75

Authors

Yonezawa, K. (deposition date: 2016-09-16, release date: 2017-08-30, Last modification date: 2023-11-08)

Primary citation

Yonezawa, K.,Shimizu, N.,Kurihara, K.,Yamazaki, Y.,Kamikubo, H.,Kataoka, M.
Neutron crystallography of photoactive yellow protein reveals unusual protonation state of Arg52 in the crystal
Sci Rep, 7:9361-9361, 2017

PubMed Abstract: Because of its high pK, arginine (Arg) is believed to be protonated even in the hydrophobic environment of the protein interior. However, our neutron crystallographic structure of photoactive yellow protein, a light sensor, demonstrated that Arg52 adopts an electrically neutral form. We also showed that the hydrogen bond between the chromophore and Glu46 is a so-called low barrier hydrogen bond (LBHB). Because both the neutral Arg and LBHB are unusual in proteins, these observations remain controversial. To validate our findings, we carried out neutron crystallographic analysis of the E46Q mutant of PYP. The resultant structure revealed that the proportion of the cationic form is higher in E46Q than in WT, although the cationic and neutral forms of Arg52 coexist in E46Q. These observations were confirmed by the occupancy of the deuterium atom bound to the N atom combined with an alternative conformation of the ND group comprising sp hybridisation. Based on these results, we propose that the formation of the LBHB decreases the proton affinity of Arg52, stabilizing the neutral form in the crystal.

PubMed: 28839266
DOI: 10.1038/s41598-017-09718-9

Experimental method

NEUTRON DIFFRACTION (1.493 Å)