10 research outputs found
The chess board pattern in the genetic code table.
<p>When all SSN and WWN codon boxes are left out, a chess board pattern emerges (see text). In this representation it can immediately be seen that mixed SW/WS codon boxes with a middle-Y (U or C) are fourfold degenerate codon boxes, while mixed SW/WS codon boxes with a middle-R (A or G) are split codon boxes.</p
Start of the genetic code with a single tRNA encoding a single amino acid (tentatively selected as Gly).
<p>In the left hand panel the codons are indicated which are in efficient and unambiguous use. In the right hand panel the anticodons are indicated which perform the efficient and unambiguous decoding of these codons. The same division between left hand panel and right hand panel is used in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158342#pone.0158342.g003" target="_blank">3</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158342#pone.0158342.g006" target="_blank">6</a>.</p
The tRNA set of present-day Archaea.
<p>The asterisks indicate anticodon first position modifications which are necessary to unambiguously read the respective codon box. The only difference with the proposed tRNA set of LUCA is the presence of the codon AUA in this codon repertoire (indicated by coloring with blue).</p
Summary of the proposed three stages in the evolutionary development of the tRNA set in the standard genetic code.
<p>Summary of the proposed three stages in the evolutionary development of the tRNA set in the standard genetic code.</p
The proposed tRNA set of LUCA.
<p>The asterisks indicate anticodon first position modifications which are necessary to unambiguously read the respective codon box. At this stage there already is a distinction between initiator methionine and elongator methionine. Red, yellow, green, and blue are used to indicate the codons where changes happened compared to the situation in the previous figure.</p
Active Site Structure of Photoactive Yellow Protein with a Locked Chromophore Analogue Revealed by Near-Infrared Raman Optical Activity
Many biological cofactors, such as
light-absorbing chromophores
in photoreceptors, are intrinsically planar molecules. A protein environment,
however, causes structural distortions of the cofactor, and such structural
changes can lead to a modulation of chemical properties of the cofactor
to maximize its biological activity. Here, we investigate the active
site structure of photoactive yellow protein (PYP), a blue light photoreceptor
that contains a <i>p</i>-coumaric acid (<i>p</i>CA) chromophore, by a near-infrared excited Raman optical activity
(ROA). Specifically, we measured the ROA spectra of PYP, whose chromophore
is replaced with a locked <i>p</i>CA analogue. Furthermore,
we show that a spectral analysis based on quantum mechanical/molecular
mechanical (QM/MM) calculations of the whole protein molecule is useful
to obtain structural information from the observed ROA spectra. The
use of the near-infrared ROA combined with QM/MM calculations is a
novel and generally applicable spectroscopic tool to study the chromophore
distortions within a protein environment
Spectroscopic Validation of Crystallographic Structures of a Protein Active Site by Chiroptical Spectroscopy
Out-of-plane distortions of a cofactor molecule in a
protein active
site are functionally important, and in photoreceptors, it has been
proposed that they are crucial for spectral tuning and energy storage
in photocycle intermediates. However, these subtle structural features
are often beyond the grasp of structural biology. This issue is strikingly
exemplified by photoactive yellow protein: its 14 independently determined
crystal structures exhibit considerable differences in the dihedral
angles defining the chromophore geometry, even though most of these
are at excellent resolution. Here we developed a strategy to verify
cofactor distortions in crystal structures by using quantum chemical
calculations and chiroptical spectroscopy, particularly Raman optical
activity and electronic circular dichroism spectroscopies. Based on
this approach, we identify seven crystal structures with the chromophore
geometries inconsistent with the experimentally observed data. The
strategy implemented here promises to be widely applicable to uncovering
cofactor distortions at active sites and to studies of reaction intermediates
Noncanonical Photocycle Initiation Dynamics of the Photoactive Yellow Protein (PYP) Domain of the PYP-Phytochrome-Related (Ppr) Photoreceptor
The
photoactive yellow protein (PYP) from <i>Halorhodospira
halophila</i> (Hhal) is a bacterial photoreceptor and model system
for exploring functional protein dynamics. We report ultrafast spectroscopy
experiments that probe photocycle initiation dynamics in the PYP domain
from the multidomain PYP-phytochrome-related photoreceptor from <i>Rhodospirillum centenum</i> (Rcen). As with Hhal PYP, Rcen PYP
exhibits similar excited-state dynamics; in contrast, Rcen PYP exhibits
altered photoproduct ground-state dynamics in which the primary I<sub>0</sub> intermediate as observed in Hhal PYP is absent. This property
is attributed to a tighter, more sterically constrained binding pocket
around the <i>p</i>-coumaric acid chromophore due to a change
in the Rcen PYP protein structure that places Phe98 instead of Met100
in contact with the chromophore. Hence, the I<sub>0</sub> state is
not a necessary step for the initiation of productive PYP photocycles
and the ubiquitously studied Hhal PYP may not be representative of
the broader PYP family of photodynamics
Subpicosecond Excited-State Proton Transfer Preceding Isomerization During the Photorecovery of Photoactive Yellow Protein
The ultrafast excited-state dynamics underlying the receptor state photorecovery is resolved in the M100A mutant of the photoactive yellow protein (PYP) from Halorhodospira halophila. The M100A PYP mutant, with its distinctly slower photocycle than wt PYP, allows isolation of the pB signaling state for study of the photodynamics of the protonated chromophore <i>cis-p</i>-coumaric acid. Transient absorption signals indicate a subpicosecond excited-state proton-transfer reaction in the pB state that results in chromophore deprotonation prior to the cis−trans isomerization required in the photorecovery dynamics of the pG state. Two terminal photoproducts are observed, a blue-absorbing species presumed to be deprotonated <i>trans-p</i>-coumaric acid and an ultraviolet-absorbing protonated photoproduct. These two photoproducts are hypothesized to originate from an equilibrium of open and closed folded forms of the signaling state, I<sub>2</sub> and I<sub>2</sub>′
Excitation-Wavelength-Dependent Photocycle Initiation Dynamics Resolve Heterogeneity in the Photoactive Yellow Protein from <i>Halorhodospira halophila</i>
Photoactive
yellow proteins (PYPs) make up a diverse class of blue-light-absorbing
bacterial photoreceptors. Electronic excitation of the <i>p</i>-coumaric acid chromophore covalently bound within PYP results in
triphasic quenching kinetics; however, the molecular basis of this
behavior remains unresolved. Here we explore this question by examining
the excitation-wavelength dependence of the photodynamics of the PYP
from <i>Halorhodospira halophila</i> via a combined experimental
and computational approach. The fluorescence quantum yield, steady-state
fluorescence emission maximum, and cryotrapping spectra are demonstrated
to depend on excitation wavelength. We also compare the femtosecond
photodynamics in PYP at two excitation wavelengths (435 and 475 nm)
with a dual-excitation-wavelength-interleaved pump–probe technique.
Multicompartment global analysis of these data demonstrates that the
excited-state photochemistry of PYP depends subtly, but convincingly,
on excitation wavelength with similar kinetics with distinctly different
spectral features, including a shifted ground-state beach and altered
stimulated emission oscillator strengths and peak positions. Three
models involving multiple excited states, vibrationally enhanced barrier
crossing, and inhomogeneity are proposed to interpret the observed
excitation-wavelength dependence of the data. Conformational heterogeneity
was identified as the most probable model, which was supported with
molecular mechanics simulations that identified two levels of inhomogeneity
involving the orientation of the R52 residue and different hydrogen
bonding networks with the <i>p</i>-coumaric acid chromophore.
Quantum calculations were used to confirm that these inhomogeneities
track to altered spectral properties consistent with the experimental
results