540
Sensory rhodopsin II: functional insights from structure
John L Spudich* and Hartmut Luecke†
Atomic resolution structures of a sensory rhodopsin phototaxis
receptor in haloarchaea (the first sensory member of the
widespread microbial rhodopsin family) have yielded insights
into the interaction face with its membrane-embedded
transducer and into the mechanism of spectral tuning.
Spectral differences between sensory rhodopsin and the
light-driven proton pump bacteriorhodopsin depend largely
upon the repositioning of a conserved arginine residue in the
chromophore-binding pocket. Information derived from the
structures, combined with biophysical and biochemical
analysis, has established a model for receptor activation and
signal relay, in which light-induced helix tilting in the receptor
is transmitted to the transducer by lateral transmembrane
helix–helix interactions.
Addresses
*Department of Biochemistry and Molecular Biology, Department of
Microbiology and Molecular Genetics, and Center for Membrane Biology,
University of Texas Medical School, Houston, Texas 77030, USA;
e-mail: John.L.Spudich@uth.tmc.edu
† Department of Molecular Biology and Biochemistry, University of
California, Irvine, California 92697, USA; e-mail: hudel@uci.edu
Current Opinion in Structural Biology 2002, 12:540–546
0959-440X/02/$ — see front matter
© 2002 Elsevier Science Ltd. All rights reserved.
Abbreviations
BR bacteriorhodopsin
λmax wavelength of maximum absorption
HR halorhodopsin
Hs Halobacterium salinarum
Np Natronobacterium pharaonis
PDB Protein Data Bank
rmsd root mean squared deviation
SDSL site-directed spin-labeling
SR sensory rhodopsin
TM transmembrane segment
Introduction
Microbial rhodopsins are a family of photoactive, seven-
transmembrane helix, retinylidene proteins found in
phylogenetically diverse microorganisms, including halo-
archaea, proteobacteria, cyanobacteria, fungi and algae [1–3].
Several rhodopsins, such as bacteriorhodopsin (BR) and
halorhodopsin (HR) in haloarchaea, and proteorhodopsin
in marine bacteria, are light-driven ion pumps, whereas
others, such as sensory rhodopsins I and II (SRI and SRII)
in haloarchaea, Chlamydomonas rhodopsins CSRA and
CSRB, and Anabaena rhodopsin (K-H Jung, VD Trivedi,
JL Spudich, unpublished data), are photosensory receptors
that are spectrally tuned throughout the visible spectrum
to relay information to the cell regarding the intensity and
color of light in the environment.
SRI and SRII are sensors for phototaxis in Halobacterium
salinarum and related halophilic archaea [4]. SRI mediates
attractant motility responses to the green/orange wavelengths
used by the ion pumps BR and HR, whereas SRII mediates
blue light avoidance responses. SRI also mediates repellent
responses to damaging near-UV light by a color-sensitive
two-photon mechanism, so that cells migrate toward
green/orange regions but not into full spectrum solar
radiation containing damaging short wavelength photons.
The SRI and SRII proteins are subunits of multicom-
ponent signaling complexes and relay signals by
protein–protein interactions to the integral membrane
transducer proteins (HtrI and HtrII, respectively) which
control a cytoplasmic phosphorylation pathway that
modulates the cell’s motility apparatus.
Among the archaeal sensory rhodopsins, the SRII homolog
in the alkalophilic haloarchaeon Natronobacterium pharaonis
(NpSRII, also called ppR for pharaonis phoborhodopsin)
has the most robust properties and is able to function
during photosignaling in N. pharaonis [5], H. salinarum [6]
and Escherichia coli [7••]. NpSRII remains stable both in the
dark and under prolonged illumination, and it retains its
native absorption spectrum and photocycle in different
membranes, in a range of detergents, in salt concentrations
from 25 mM to 4 M, and over a broad range of pH.
These exceptional properties have made NpSRII the
prototypical sensory rhodopsin for crystallography and
structure/function studies.
SRI and HtrI function in a 2:2 complex in their native
H. salinarum membranes [8••]. NpHtrII forms homodimers,
as shown by electron paramagnetic resonance dipolar
interactions between spin-labeled monomers in proteo-
liposomes [9••] and by rapid disulfide cross-linking in
H. salinarum membranes [10••]. A molar ratio of 1:1 has
been found for NpSRII and NpHtrII in proteoliposomes
[9••] and in nondenaturing detergents that maintain the
complex [11,12••]; therefore, NpSRII and NpHtrII also
appear to form 2:2 molecular complexes. Deletion and
chimera experiments with the H. salinarum transducers
revealed that the interaction specificities of SRI with HtrI
and SRII with HtrII are determined by the transmembrane
helices of the Htr subunits, indicating that interaction
occurs within or near the membrane [13]. This is confirmed
for NpHtrII as well, because a truncated form that
lacks most of the cytoplasmic domain interacts with
NpSRII in vitro [9••,11,12••] and is functional in vivo [7••].
Our current view of the overall composition of the
NpSRII–HtrII signaling complex is depicted in Figure 1.
Insights from the crystal structures
A projection structure obtained from 2D crystals of NpSRII
by cryoelectron microscopy [14] showed that NpSRII
was indistinguishable from BR at low resolution (6.9 Å).
Hence, understanding the different properties of these
 Sensory rhodopsin II Spudich and Luecke 541

two proteins clearly requires the comparison of their Figure 1

structures at atomic resolution. Two recent atomic resolution
X-ray structures of NpSRII, obtained from crystals grown SR
in cubic lipid phase [15], show a high level of agreement
(rmsd of backbone atoms 0.47 Å). One structure was
obtained by expression of the N. pharaonis receptor in the
related haloarchaeon H. salinarum, followed by extraction
with octyl glucoside, and using KCl at pH 5.3 to induce
crystallization ([16••]; PDB code 1JGJ). The second
study used expression of NpSII in E. coli in the presence
of 10 µM all-trans retinal, followed by extraction with
dodecyl maltoside, and growth of crystals at the lower pH
of 4.6 in the presence of NaCl ([17••]; PDB code 1H68).
Cytoplasm
Both crystallization protocols required the addition of
polar H. salinarum lipids and yielded nearly isomorphous
crystals, displaying the same space group and similar unit
cell dimensions. The pH difference might explain why
the former structure does not contain ordered chloride, Htr
whereas the latter contains a partially occupied putative
chloride ion (Figure 2).

Recently, photoisomerization-induced changes in the CheW

microenvironment of the retinal have been characterized
by low temperature light/dark difference Fourier transform
infrared (FTIR) spectroscopy [18] and by X-ray analysis
of illuminated crystals at 100 K [19]. The structural
rearrangements in the retinal pocket are similar to those
studied in the early BR photocycle intermediate K [20],
but with more extensive movement of the chromophore
and, in particular, more movement of its β-ionone ring
than in BR. However, because of limited resolution and
partial occupancies, neither the BR nor the NpSRII
intermediate study provided information about the
Schiff base configuration. Other biophysical methods
over the past two years have generated information
regarding NpSRII photophysical properties and its
photochemical reaction cycle [21–32].
The X-ray structures provide insight into two important
questions regarding the receptors properties: ‘What is
the mechanism of spectral tuning in NpSRII?’ and ‘Which
face of the NpSRII molecule interacts with its cognate
NpHtrII transducer?’
What is the mechanism of spectral tuning in NpSRII?
NpSRII absorbs maximally (λmax) at 498 nm, which is
≥70 nm shifted to shorter wavelengths than those of BR,
HR and HsSRI (568 nm, 576 nm and 587 nm, respectively).
The crystal structures mentioned previously provide
insight into the retinal–protein interactions that are
responsible for this difference. Free in solution, a proto-
nated retinylidene Schiff base exhibits an absorption
maximum of 440 nm. A shift from this value to longer
wavelengths caused by interactions of the chromophore
with protein residues is called the ‘opsin shift’ [33].
Nearby charges and polar residues, as well as strain on the
extended conjugated electronic system of the retinal,
influence the energy difference between the electronic
CheA
P P
CheY CheY
Flagellar
motor
Current Opinion in Structural Biology
Schematic drawing of the repellent phototaxis signaling system in
N. pharaonis based on experiments reviewed in the text and
well-established features of the partially homologous bacterial
chemotaxis system [4,49]. NpSRII receptors are drawn as interacting
in a symmetrical 2:2 complex with the transmembrane helices of the
NpHtrII dimer, which is bound to the histidine kinase CheA by a small
protein (CheW). Photoactivation of NpSRII activates CheA kinase
activity, which in turn phosphorylates the flagellar response regulator
CheY. Phospho-CheY binds to a flagellar motor switch that induces a
swimming reversal by the cell, leading to repulsion from NpSRII-absorbed
light (λmax 498 nm). The methylation-dependent adaptation components
are not shown.
ground and excited states, thereby contributing to the
opsin shift [33,34]. However, the substitution of residues in
the NpSRII retinal-binding pocket with the corresponding
BR residues recovered only 30% of the relative opsin shift
in the absorption maximum compared to that of BR [35].
Even a ten-residue mutant, in which all NpSRII residues
within 5 Å of the retinal were converted to their BR
equivalents, resulted in only 44% recovery compared to
BR [36•]. Thus, over half of the difference in opsin
shifts between NpSRII and BR is not accounted for by
542 Membranes

Figure 2

Stereo view of the region of the putative

chloride in NpSRII (PDB code 1H68)
described by Royant et al. [17••]. After
adding the missing atoms for residues Asp28,
Ser31, Glu33, Arg35, Glu97 and Leu219 for
PDB entry 1H68 [17••] using the Swiss-PDB
Viewer version 3.7b2, alignment with
PDB entry 1JGJ [16••] using the ‘Iterative
Magic Fit’ procedure resulted in an rmsd for
backbone atoms of 0.47 Å. The largest
mainchain deviations are at the carbonyl of
Val63 near the turn of the BC loop (Ala64
is a Ramachandran outlier in 1H68), in the
region from Arg27 to Ser31 (Ala29 is a
Ramachandran outlier in 1H68), and at
Leu187. Further inspection of the 1H68
PDB entry reveals that its putative chloride
(CL500; green circle) is modeled with 50% of Asp193: in the 1H68 structure, the only possible when the carboxylate is
occupancy, occupying the same space as a carboxyl group is rotated about χ1, resulting protonated, it seems likely that the 1H68
50% occupied water molecule (Wat-501). in a movement of 1.0 Å away from the entry represents a structure where Asp193 is
In PDB entry 1JGJ there is a fully-occupied guanidinium of Arg72. This movement brings protonated, possibly due to the lower pH
water 0.6 Å from this site (Wat-601); however, the OD2 atom of Asp193 to within 2.91 Å of (pH4.6 versus pH5.3 for 1JGJ). It is
in this study no indication of an ordered chloride the mainchain carbonyl oxygen of Ile177, conceivable that only the loss of the negative
was noted. A probably related structural suggesting a hydrogen bond. As a hydrogen charge of Asp193 at low pH would enable a
difference in this region exists for the sidechain bond between a carboxylate and a carbonyl is chloride ion to bind in this region.

these ten local mutations, hinting at the involvement of
significant long-range effectors.
The X-ray structures reveal that the residues in the NpSRII
retinal-binding pocket, including the nearby negatively
charged Asp75 and Asp201 (Figure 3) do not significantly
change their position relative to the retinal. However, about
10 Å from the Schiff base the positively charged guanidinium
moiety of residue Arg72 increases its distance from the
protonated Schiff base relative to that of Arg82 in BR by
over 1 Å, and was subsequently shown by molecular orbital
theory calculations to be the single most important
contributor to the λmax difference [37••], although a slight
difference in the position of Asp75 was emphasized in
another theoretical study [38] based on QM/MM (quantum
mechanical/molecular mechanical) optimization of a pre-
liminarily refined version of the structure later published
[17••]. Thus, somewhat unexpectedly, the reorientation of
a conserved and distant residue has a larger effect on the
electronic states of the retinal chromophore than nearby
mutations. A smaller contribution is made by the hydroxyl
group of Thr205 (Ala215 in BR) at the π-bulge [39•], which
preferentially stabilizes the ground state. A further factor
appears to be the increased space in the β-ionone binding
pocket, where smaller residues in NpSRII (Gly130,
Ala131 versus Ser141, Thr142 in BR) allow a slightly more
extended (unbent) polyene chain in comparison to BR.
The relatively pronounced vibrational fine structure in the
absorption spectra of NpSRII proteins may also be due in
part to the repositioning of Arg72, although the origins of
the fine structure could not be identified with certainty
from the molecular orbital theory calculations [37••].
Which face of the NpSRII molecule interacts with its
cognate NpHtrII transducer?
The structures of NpSRII revealed that a tyrosine residue
protrudes from the middle of the lipid-facing surface of
helix G (Figure 4). The tyrosine phenol group is too long
and rigid to allow the formation of a hydrogen bond to the
mainchain carbonyls of its own helix, unlike threonine or
serine hydroxyls. To avoid an unpaired polar hydroxyl
in the middle of the bilayer in the crystals, Tyr199 forms
an intermolecular hydrogen bond to a carbonyl of a trans-
membrane helix from a neighboring molecule in the same
bilayer [16••]. Tyr199 is conserved in the three known
SRII sequences, whereas this residue is a phenylalanine in
the two known SRI sequences [4]. The need for Tyr199
to hydrogen bond to an adjacent protein makes Tyr199 an
excellent candidate for transducer binding in the
SRII–HtrII complex in N. pharaonis membranes. The
hydrophobic surface of NpSRII displays three distinct
faces: face I is formed by helices F, G and A; face II by
helices A (cytoplasmic half), B, C, D and E (extracellular
half); and face III by helices E and F. Tyr199 is located
prominently in the middle of face I — the face that also
contains both helices known to undergo large motions
during the BR photocycle [40,41]. It is therefore likely that
face I provides the majority of the interaction with the
photosignal transducer NpHtrII.
A recent in vitro binding study [12••] reported that
mutation of Tyr199 to phenylalanine reduced the binding
affinity of purified NpSRII D75N mutant to a truncated
fragment of NpHtrII by 10-fold, confirming the participation
of this face I residue in binding the transducer. Other
residues on the outer face of helix G also show clear
 Sensory rhodopsin II Spudich and Luecke 543

Figure 3

Stereo figure comparing the photoreceptor

NpSRII (PDB code 1JGJ) with the ion pump
BR (1C3W). The coordinates were aligned
using the ‘Iterative Magic Fit’ procedure of the
Swiss-PDBViewer, yielding an rmsd of 0.96 Å
for 816 mainchain atoms (BR is shown in
cerise, NpSRII in conventional atom colors).
Overall, the structures are very similar; however, Asp201 Asp201

the distance from the protonated Schiff base

nitrogen (Lys205 NZ) to the arginine
guanidinium (Arg72) in the extracellular
portion of the receptor increases (to 10.5 Å
from 9.4 Å in BR), whereas other key residues
nearer to the Schiff base, including the pair of
counterions, Asp75 and Asp201, retain their
distances (Asp75: 4.42 Å versus 4.45 Å in
BR; Asp201: 4.25 Å versus 4.12 Å in BR).
For NpSRII from PDB entry 1H68 the
distances are very similar (Asp75: 4.58 Å;
Asp201: 4.11 Å).

grouping between the SRI and SRII subfamilies, possibly
contributing to transducer binding specificity. Further
evidence is provided by the site-directed spin-labeling
(SDSL) electron paramagnetic resonance (EPR) study by
Wegener et al. [9••], which reported that three amino acids
at the cytoplasmic end of face I (Ser158, Phe210, Ile211)
interact with NpHtrII (Figure 4). Consistent with face I
binding, Royant et al. [17••] suggest that a patch of positive
charges on the cytoplasmic side of face I interacts with
acidic groups predicted by the NpHtrII sequence to be on
its cytoplasmic domain [41].
The signal transfer mechanism and future
directions
The next challenges for the crystallographic analysis of
NpSRII are to obtain the structures of light-induced
photointermediates that function as signaling states in
the receptor (the long-lived M and O intermediates that
appear late in the photocycle appear to fulfill this role [4]),
and to obtain structures of the receptor–transducer
complex in the dark and in the light. Both efforts will test
and refine models for the transmission of signals from
SR to Htr proteins. A model based on studies of the
SRI–HtrI and SRII–HtrII complexes of H. salinarum
[42,43••] was derived in part from the observation that
SRI pumps protons, but only when it is free of its
transducer. The HtrI protein was found to inhibit SRI
pumping by closing a cytoplasmic ion-conducting channel
during its photocycle. A tilting of helices, primarily
helix F, contributes to the opening of a cytoplasmic
channel in the latter half of the photocycle in BR [40].
The unified mechanism model is based on the proposal

Figure 4

Putative face of interaction between the

NpSRII photoreceptor and its cognate
transducer, NpHtrII. The NpSRII surface
(cytoplasmic side at top) is colored
according to amino acid type (red: negatively
charged; blue: positively charged; yellow:
polar; graey: hydrophobic). Residue Tyr199
is situated in the middle of the bilayer (whose
hydrophobic portion lies between the white
horizontal lines) near a π-bulge on the
outside of helix G one-and-a-half helical
turns from the lysine (Lys205) to which the
retinal is attached. Also depicted in purple are
residues implicated in transducer interaction
by SDSL EPR (Ser158/Phe210/Ile211,
[9••]). A patch of three positively charged
residues (Lys157, Arg162, Arg164, shown
in blue around Ser158) has also been
proposed to constitute a transducer binding
site, based on the absence of such a patch
in BR and HR [17••].
544 Membranes
that ion transport and sensory signaling use the same
retinal-driven protein structural changes. The key feature
of the model is that light-induced disruption of the salt
bridge between the protonated Schiff base on helix G
and the aspartyl counterion on helix C triggers tilting of
helix F, to which the Htr transmembrane helices are
coupled. Supporting a unified molecular mechanism,
substitution of the Schiff base counterion Asp85 with
asparagine induces helix F tilting in the dark in BR [44];
the corresponding substitution in HsSRII partially
activates the receptor in the dark [45].
Recently, several results obtained with NpSRII confirm
the predictions of the unified mechanism model. SDSL
demonstrated a transient light-induced tilting of helix F
in NpSRII [46•], similar to that which occurs in BR.
Electrophysiological and ion-flux studies of HsSRI and
NpSRII confirmed that HsHtrI blocks HsSRI transport
[47•] and, furthermore, that light-driven transport by
NpSRII, which was more difficult to demonstrate, is also
blocked by NpHtrII interaction [47•,48•]. These results
show that both HsSRI and NpSRII receptors contain,
under appropriate conditions, open cytoplasmic channels
during their photocycles, and strongly suggest that their
respective transducers are coupled to channel opening, a
basic tenet of the model.
An additional prediction is that, because SR helix tilting
in the unified mechanism model is transmitted to the
Htr protein by direct helix–helix contacts, alterations in
structure must occur in the Htr transmembrane domains
between the receptor interaction sites and the cyto-
plasmic domain of the transducer, where the activity of
the bound histidine kinase is controlled. Such structural
alterations have been detected as light-induced changes
in interactions between spin labels introduced into the
NpHtrII transmembrane helices [9••] and as changes of
disulfide bond formation rates between engineered
cysteines [10••]. In both studies, the data support that
the second transmembrane segment (TM2) of NpHtrII
is more conformationally active than TM1. The authors
of the SDSL study further suggest a signal transfer
mechanism in which TM2 undergoes a rotary motion in
response to the tilting of helix F in the photoactivated
receptor [9••].
Conclusions
These early glimpses of the receptor–transducer signaling
mechanism have helped to guide efforts towards obtaining
an atomic resolution moving picture of NpSRII photo-
activation, and signal relay to NpHtrII. Such efforts in the
future will contribute an example of dynamic protein–protein
interaction within a membrane, a fundamental process about
which very little is known.
Acknowledgements
The authors’ work referred to in this review was supported by grants from
the National Institutes of Health (R01-GM27750 to JLS and R01-GM56445
to HL), and a Robert A Welch Foundation Award to JLS.
References and recommended reading
Papers of particular interest, published within the annual period of review,
have been highlighted as:
• of special interest
•• of outstanding interest
1. Spudich JL, Yang C-H, Jung K-H, Spudich EN: Retinylidene Proteins:
structures and functions from archaea to humans. Ann Rev Cell
Dev Biol 2000, 16:365-392.
2. Béjà O, Spudich EN, Spudich JL, Leclerc M, DeLong EF:
Proteorhodopsin phototrophy in the ocean. Nature 2001,
411:786-789.
3. Sineshchekov OA, Jung K-H, Spudich JL: Two rhodopsins mediate
phototaxis to low and high intensity light in Chlamydomonas
reinhardtii. Proc Natl Acad Sci USA 2002 (published online
11 June 2002) DOI:10.1073/pnas.122243399.
4. Hoff WD, Jung K-H, Spudich JL: Molecular mechanism of
photosignaling by archaeal sensory rhodopsins. Annu Rev
Biophys Biomol Struct 1997, 26:223-258.
5. Scharf B, Wolff EK: Phototactic behaviour of the archaebacterial
Natronobacterium pharaonis. FEBS Lett 1994, 340:114-116.
6. Luttenberg B, Wolff EK, Engelhard M: Heterologous coexpression
of the blue light receptor psRII and its transducer pHtrII from
Natronobacterium pharaonis in the Halobacterium salinarium
strain Pho81/w restores negative phototaxis. FEBS Lett 1998,
426:117-120.
7. Jung K-H, Spudich EN, Trivedi VD, Spudich JL: An archaeal
•• photosignal-transducing module mediates phototaxis in
Escherichia coli. J Bacteriol 2001, 183:6365-6371.
Fusion-chimera proteins consisting of three genetically fused parts (NpSRII,
the N-terminal portion of NpHtrII that contains two transmembrane segments,
and cytoplasmic domains of eubacterial chemotaxis receptors) were shown
to mediate retinal-dependent phototaxis responses in E. coli. The results
establish that the nine-helix membrane portion of the receptor–transducer
complex is a modular functional unit able to signal in heterologous membranes.
The authors conclude that NpHtrII interacts physically and functionally
with NpSRII via helix–helix contacts between their transmembrane helices,
as was suggested from chimera experiments that showed that SR receptor
interaction specificity was encoded in the Htr transducer transmembrane
helices [13].
8. Chen X, Spudich JL: Demonstration of 2:2 stoichiometry in the
•• SRI-HtrI signaling complex in Halobacterium salinarum
membranes by gene-fusion analysis. Biochemistry 2002,
41:3891-3896.
SRI and HtrI were forced into a 1:1 molar ratio by gene fusion, and
dimerization of the HtrI portion resulted in a 2:2 complex active in phototaxis
signaling. Therefore, the 2:2 composition of SR receptors and Htr transducers
observed for reconstituted purified proteins in vitro [9••,11] occurs also in
functional complexes in natural membranes.
9. Wegener AA, Klare JP, Engelhard M, Steinhoff HJ: Structural insights
•• into the early steps of receptor-transducer signal transfer in
archaeal phototaxis. EMBO J 2001, 20:5312-5319.
EPR with SDSL was used to measure distances between spin-labels placed
at sites on NpSRII and NpHtrII in the dark and in the photoactivated state.
Light/dark difference EPR spectra as well as time-resolved changes in
selected positions on the receptor and transducer were recorded. The
results support the unified mechanism and, furthermore, provide insight
into the molecular details of the signal relay from receptor to transducer.
Light-insensitive dipolar interactions show that helix G of the receptor and
the two TM1 helices of the transducer appear to be fixed in position, whereas
helix F of the receptor and the two TM2 helices undergo substantial light-
induced displacements. The distance changes are consistent with a
clockwise rotation of TM2 in response to the helix F tilt. A piston-like motion
of TM2, such as that which has been detected in bacterial chemotaxis
receptors [49], is not excluded by the data. Of particular interest, the
time-resolved data reveal a time delay of a factor of four between the reset
motion of helix F and that of TM2. The transient persistence of conformational
change in the transducer after the receptor returns to the unstimulated
state would amplify the signal, thereby increasing light-sensitivity of the
signaling complex.
10. Yang CS, Spudich JL: Light-induced structural changes occur in
•• the transmembrane helices of the Natronobacterium pharaonis
HtrII transducer. Biochemistry 2001, 40:14207-14214.
Oxidative disulfide cross-linking efficiencies of monocysteine mutants of
NpHtrII were measured with or without photoactivation of NpSRII.
Conformationally active regions in TM2 of the transducer and in a face along
the length of TM2, which becomes more available for TM2–TM2′ cross-
linking upon receptor photoactivation are identified. Of note is that these

data, obtained from functional complexes in membranes, are fully in agreement
with the results from EPR spectroscopy of the complex obtained with
reconstituted purified receptor and truncated transducer [9••]. Both studies
report substantial conformational activity in TM2 and that Ala80 and Thr81
are active residues. The data also establish that one residue in TM2, Gly83,
is critical for maintaining the proper conformation of NpHtrII for signal relay from
the photoactivated receptor to the kinase-binding region of the transducer.
11. Sudo Y, Iwamoto M, Shimono K, Kamo N: Pharaonis phoborhodopsin
binds to its cognate truncated transducer even in the presence of
a detergent with a 1:1 stoichiometry. Photochem Photobiol 2001,
74:489-494.
12. 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, 83:427-432.
The authors provide direct evidence that Tyr199 participates in the binding
interaction of NpSRII with the transducer NpHtrII. This paper, together with
[11] also establishes a rapid quantitative assay for binding of purified
solubilized NpSRII and NpHtrII, based on easily measured effects of the
transducer on the receptor photocycle.
13. Zhang X-N, Zhu J, Spudich JL: The specificity of interaction of
archaeal transducers with their cognate sensory rhodopsins is
determined by their transmembrane helices. Proc Natl Acad Sci
USA 1999, 273:19722-19728.
14. Kunji ERS, Spudich EN, Grisshammer R, Henderson R, Spudich JL:
Electron crystallographic analysis of two-dimensional crystals of
sensory rhodopsin II: A 6.9 Å projection structure. J Mol Biol 2001,
308:279-293.
15. Landau EM, Rosenbusch JP: Lipidic cubic phases: a novel concept
for the crystallization of membrane proteins. Proc Natl Acad Sci
USA 1996, 93:14532-14535.
16. Luecke H, Schobert B, Lanyi JK, Spudich EN, Spudich JL: Crystal
•• structure of sensory rhodopsin II at 2.4 Å: insights into color
tuning and transducer interaction. Science 2001, 293:1499-1503.
The first high-resolution structure of NpSRII (PDB code 1JGJ), which, compared
to the structure of BR, revealed relatively minor changes near the chromophore
with slightly less strain on its conjugated π system, but a 1 Å repositioning of
the more distant positive charge of Arg72. The cytoplasmic side is significantly
more hydrophobic than that of BR, and bilayer-exposed face I was proposed
to interact with the cognate transducer, NpHtrII.
17. Royant A, Nollert P, Edman K, Neutze R, Landau EM, Pebay-Peyroula E,
•• Navarro J: X-ray structure of sensory rhodopsin II at 2.1 A
resolution. Proc Natl Acad Sci USA 2001, 98:10131-10136.
An independent high-resolution NpSRII structure (PDB code 1H68) that
agrees well with 1JGJ [16••] with the exception of an additional partially
occupied putative chloride ion near Arg72.
18. Kandori H, Shimono K, Sudo Y, Iwamoto M, Shichida Y, Kamo N: Structural
changes of pharaonis phoborhodopsin upon photoisomerization of
the retinal chromophore: infrared spectral comparison with
bacteriorhodopsin. Biochemistry 2001, 40:9238-9246.
19. Edman K, Royant A, Nollert P, Maxwell CA, Pebay-Peyroula E,
Navarro J, Neutze R, Landau EM: Early structural rearrangements in
the photocycle of an integral membrane sensory receptor.
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