A Long-lived M-like State of Phoborhodopsin That Mimics the Active State

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2008 Apr 1

PMID 18375514

Citations 4

Authors

Yuki Sudo

Tatsuya Nishihori

Masayuki Iwamoto

Kazumi Shimono

Chojiro Kojima

Naoki Kamo

Affiliations

Soon will be listed here.

Abstract

Pharaonis phoborhodopsin (ppR, also called pharaonis sensory rhodopsin II) is a seven transmembrane helical retinal protein. ppR forms a signaling complex with pharaonis Halobacterial transducer II (pHtrII) in the membrane that transmits a light signal to the sensory system in the cytoplasm. The M-state during the photocycle of ppR (lambda(max) = 386 nm) is one of the active (signaling) intermediates. However, progress in characterizing the M-state at physiological temperature has been slow because its lifetime is very short (decay half-time is approximately 1 s). In this study, we identify a highly stable photoproduct that can be trapped at room temperature in buffer solution containing n-octyl-beta-d-glucoside, with a decay half-time and an absorption maximum of approximately 2 h and 386 nm, respectively. HPLC analysis revealed that this stable photoproduct contains 13-cis-retinal as a chromophore. Previously, we reported that water-soluble hydroxylamine reacts selectively with the M-state, and we found that this stable photoproduct also reacts selectively with that reagent. These results suggest that the physical properties of the stable photoproduct (named the M-like state) are very similar with the M-state during the photocycle. By utilizing the high stability of the M-like state, we analyzed interactions of the M-like state and directly estimated the pK(a) value of the Schiff base in the M-like state. These results suggest that the dissociation constant of the ppR(M-like)/pHtrII complex greatly increases (to 5 muM) as the pK(a) value greatly decreases (from 12 to 1.5). The proton transfer reaction of ppR from the cytoplasmic to the extracellular side is proposed to be caused by this change in pK(a).

Citing Articles

The Blue-Green Sensory Rhodopsin SRM from Haloarcula marismortui Attenuates Both Phototactic Responses Mediated by Sensory Rhodopsin I and II in Halobacterium salinarum.

Chen J, Lin Y, Fu H, Yang C Sci Rep. 2019; 9(1):5672.

PMID: 30952934 PMC: 6450946. DOI: 10.1038/s41598-019-42193-y.

Phototactic and chemotactic signal transduction by transmembrane receptors and transducers in microorganisms.

Suzuki D, Irieda H, Homma M, Kawagishi I, Sudo Y Sensors (Basel). 2012; 10(4):4010-39.

PMID: 22319339 PMC: 3274258. DOI: 10.3390/s100404010.

An active photoreceptor intermediate revealed by in situ photoirradiated solid-state NMR spectroscopy.

Tomonaga Y, Hidaka T, Kawamura I, Nishio T, Ohsawa K, Okitsu T Biophys J. 2011; 101(10):L50-2.

PMID: 22098758 PMC: 3218342. DOI: 10.1016/j.bpj.2011.10.022.

Protein-protein interaction changes in an archaeal light-signal transduction.

Kandori H, Sudo Y, Furutani Y J Biomed Biotechnol. 2010; 2010:424760.

PMID: 20671933 PMC: 2910557. DOI: 10.1155/2010/424760.

References

Radzwill N, Gerwert K, Steinhoff H . Time-resolved detection of transient movement of helices F and G in doubly spin-labeled bacteriorhodopsin. Biophys J. 2001; 80(6):2856-66. PMC: 1301470. DOI: 10.1016/S0006-3495(01)76252-2. View

Sudo Y, Iwamoto M, Shimono K, Kamo N . Role of charged residues of pharaonis phoborhodopsin (sensory rhodopsin II) in its interaction with the transducer protein. Biochemistry. 2004; 43(43):13748-54. DOI: 10.1021/bi048803c. View

Wegener A, Chizhov I, Engelhard M, Steinhoff H . Time-resolved detection of transient movement of helix F in spin-labelled pharaonis sensory rhodopsin II. J Mol Biol. 2000; 301(4):881-91. DOI: 10.1006/jmbi.2000.4008. View

Sasaki J, Kumauchi M, Hamada N, Oka T, Tokunaga F . Light-induced unfolding of photoactive yellow protein mutant M100L. Biochemistry. 2002; 41(6):1915-22. DOI: 10.1021/bi011721t. View

Shimono K, Hayashi T, Ikeura Y, Sudo Y, Iwamoto M, Kamo N . Importance of the broad regional interaction for spectral tuning in Natronobacterium pharaonis phoborhodopsin (sensory rhodopsin II). J Biol Chem. 2003; 278(26):23882-9. DOI: 10.1074/jbc.M301200200. View

Rudolph J, Oesterhelt D . Deletion analysis of the che operon in the archaeon Halobacterium salinarium. J Mol Biol. 1996; 258(4):548-54. DOI: 10.1006/jmbi.1996.0267. View

Iwamoto M, Furutani Y, Sudo Y, Shimono K, Kandori H, Kamo N . Role of Asp193 in chromophore-protein interaction of pharaonis phoborhodopsin (sensory rhodopsin II). Biophys J. 2002; 83(2):1130-5. PMC: 1302214. DOI: 10.1016/S0006-3495(02)75236-3. View

Sudo Y, Iwamoto M, Shimono K, Kamo N . Association of pharaonis phoborhodopsin with its cognate transducer decreases the photo-dependent reactivity by water-soluble reagents of azide and hydroxylamine. Biochim Biophys Acta. 2001; 1558(1):63-9. DOI: 10.1016/s0005-2736(01)00423-0. View

Furutani Y, Kamada K, Sudo Y, Shimono K, Kamo N, Kandori H . Structural changes of the complex between pharaonis phoborhodopsin and its cognate transducer upon formation of the M photointermediate. Biochemistry. 2005; 44(8):2909-15. DOI: 10.1021/bi047893i. View

10.

Sudo Y, Iwamoto M, Shimono K, Kamo N . Tyr-199 and charged residues of pharaonis Phoborhodopsin are important for the interaction with its transducer. Biophys J. 2002; 83(1):427-32. PMC: 1302158. DOI: 10.1016/S0006-3495(02)75180-1. View

11.

Klare J, Bordignon E, Engelhard M, Steinhoff H . Sensory rhodopsin II and bacteriorhodopsin: light activated helix F movement. Photochem Photobiol Sci. 2004; 3(6):543-7. DOI: 10.1039/b402656j. View

12.

Chen X, Spudich J . Demonstration of 2:2 stoichiometry in the functional SRI-HtrI signaling complex in Halobacterium membranes by gene fusion analysis. Biochemistry. 2002; 41(12):3891-6. DOI: 10.1021/bi015966h. View

13.

Imasheva E, Shimono K, Balashov S, Wang J, Zadok U, Sheves M . Formation of a long-lived photoproduct with a deprotonated Schiff base in proteorhodopsin, and its enhancement by mutation of Asp227. Biochemistry. 2005; 44(32):10828-38. DOI: 10.1021/bi050438h. View

14.

Devanathan S, Genick U, Canestrelli I, Meyer T, Cusanovich M, Getzoff E . New insights into the photocycle of Ectothiorhodospira halophila photoactive yellow protein: photorecovery of the long-lived photobleached intermediate in the Met100Ala mutant. Biochemistry. 1998; 37(33):11563-8. DOI: 10.1021/bi9803776. View

15.

Moukhametzianov R, Klare J, Efremov R, Baeken C, Goppner A, Labahn J . Development of the signal in sensory rhodopsin and its transfer to the cognate transducer. Nature. 2006; 440(7080):115-9. DOI: 10.1038/nature04520. View

16.

Shimono K, Ikeura Y, Sudo Y, Iwamoto M, Kamo N . Environment around the chromophore in pharaonis phoborhodopsin: mutation analysis of the retinal binding site. Biochim Biophys Acta. 2001; 1515(2):92-100. DOI: 10.1016/s0005-2736(01)00394-7. View

17.

Royant A, Nollert P, Edman K, Neutze R, Landau E, Pebay-Peyroula E . X-ray structure of sensory rhodopsin II at 2.1-A resolution. Proc Natl Acad Sci U S A. 2001; 98(18):10131-6. PMC: 56927. DOI: 10.1073/pnas.181203898. View

18.

Wegener A, Klare J, Engelhard M, Steinhoff H . Structural insights into the early steps of receptor-transducer signal transfer in archaeal phototaxis. EMBO J. 2001; 20(19):5312-9. PMC: 125640. DOI: 10.1093/emboj/20.19.5312. View

19.

Shimono K, Iwamoto M, Sumi M, Kamo N . Functional expression of pharaonis phoborhodopsin in Escherichia coli. FEBS Lett. 1998; 420(1):54-6. DOI: 10.1016/s0014-5793(97)01487-7. View

20.

Chizhov I, Schmies G, Seidel R, Sydor J, Luttenberg B, Engelhard M . The photophobic receptor from Natronobacterium pharaonis: temperature and pH dependencies of the photocycle of sensory rhodopsin II. Biophys J. 1998; 75(2):999-1009. PMC: 1299773. DOI: 10.1016/S0006-3495(98)77588-5. View