948 research outputs found
Extrusion of the C-terminal helix in navel orangeworm moth pheromone-binding protein (AtraPBP1) controls pheromone binding
The navel orangeworm, Amyelois transitella (Walker), is an agricultural insect pest that can be controlled by disrupting maleāfemale communication with sex pheromones, a technique known as mating disruption. Insect pheromone-binding proteins (PBPs) provide fast transport of hydrophobic pheromones through aqueous sensillar lymph and promote sensitive delivery of pheromones to receptors. Here we present a mutational analysis on a PBP from A. transitella (AtraPBP1) to evaluate how the C-terminal helix in this protein controls pheromone binding as a function of pH. Pheromone binds tightly to AtraPBP1 at neutral pH, but the binding is much weaker at pH below 5. Deletion of the entire C-terminal helix (residues 129ā142) causes more than 100-fold increase in pheromone-binding affinity at pH 5 and only a 1.5-fold increase at pH 7. A similar pH-dependent increase in pheromone binding is also seen for the H80A/H95A double mutant that promotes extrusion of the C-terminal helix by disabling salt bridges at each end of the helix. The single mutants (H80A and H95A) also exhibit pheromone binding at pH below 5, but with ā¼2-fold weaker affinity. NMR and circular dichroism data demonstrate a large overall structural change in each of these mutants at pH 4.5, indicating an extrusion of the C-terminal helix that profoundly affects the overall structure of the low pH form. Our results confirm that sequestration of the C-terminal helix at low pH as seen in the recent NMR structure may serve to block pheromone binding. We propose that extrusion of these C-terminal residues at neutral pH (or by the mutations in this study) exposes a hydrophobic cleft that promotes high affinity pheromone binding
Odorant-Binding Proteins of the Malaria Mosquito Anopheles funestus sensu stricto
Background
The mosquito Anopheles funestus is one of the major malaria vector species in sub-Saharan Africa. Olfaction is essential in guiding mosquito behaviors. Odorant-binding proteins (OBPs) are highly expressed in insect olfactory tissues and involved in the first step of odorant reception. An improved understanding of the function of malaria mosquito OBPs may contribute to identifying new attractants/repellents and assist in the development of more efficient and environmentally friendly mosquito controlling strategies.
Methodology
In this study, a large screening of over 50 ecologically significant odorant compounds led to the identification of 12 ligands that elicit significant electroantennographic (EAG) responses from An. funestus female antennae. To compare the absolute efficiency/potency of these chemicals, corrections were made for differences in volatility by determining the exact amount in a stimulus puff. Fourteen AfunOBP genes were cloned and their expression patterns were analyzed. AfunOBP1, 3, 7, 20 and 66 showed olfactory tissue specificity by reverse transcriptase PCR (RT-PCR). Quantitative real-time PCR (qRT-PCR) analysis showed that among olfactory-specific OBPs, AfunOBP1 and 3 are the most enriched OBPs in female antennae. Binding assay experiments showed that at pH 7, AfunOBP1 significantly binds to 2-undecanone, nonyl acetate, octyl acetate and 1-octen-3-ol but AfunOBP3, which shares 68% identify with AfunOBP1 at amino acid level, showed nearly no binding activity to the selected 12 EAG-active odorant compounds.
Conclusion
This work presents for the first time a study on the odorants and OBPs of the malaria vector mosquito An. funestus, which may provide insight into the An. funestus olfactory research, assist in a comparative study between major malaria mosquitoes An. gambiae and An. funestus olfactory system, and help developing new mosquito control strategies to reduce malaria transmission
1H, 15N, and 13C chemical shift assignments of the mosquito odorant binding protein-1 (CquiOBP1) bound to the mosquito oviposition pheromone
An odorant-binding protein from the Southern house mosquito, Culex pipiens quinquefasciatus (Cqui-OBP1) binds to the mosquito oviposition pheromone (MOP), 6-acetoxy-5-hexadecanolide to facilitate the transport of MOP to membrane-bound odorant receptors. We report complete NMR chemical shift assignments of Cqui-OBP1 bound to the MOP pheromone obtained at pH 7.0 and 25Ā°C (BMRB no. 16175)
Olfactory proteins mediating chemical communication in the Navel Orangeworm Moth, Amyelois transitella
Background
The navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae), is the most serious insect pest of almonds and pistachios in California for which environmentally friendly alternative methods of control ā like pheromone-based approaches ā are highly desirable. Some constituents of the sex pheromone are unstable and could be replaced with parapheromones, which may be designed on the basis of molecular interaction of pheromones and pheromone-detecting olfactory proteins.
Methodology
By analyzing extracts from olfactory and non-olfactory tissues, we identified putative olfactory proteins, obtained their N-terminal amino acid sequences by Edman degradation, and used degenerate primers to clone the corresponding cDNAs by SMART RACE. Additionally, we used degenerate primers based on conserved sequences of known proteins to fish out other candidate olfactory genes. We expressed the gene encoding a newly identified pheromone-binding protein, which was analyzed by circular dichroism, fluorescence, and nuclear magnetic resonance, and used in a binding assay to assess affinity to pheromone components.
Conclusion
We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor. Of these, AtraPBP1 is highly enriched in male antennae. Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH
Structure of an Odorant-Binding protein from the Mosquito Aedes aegypti suggests a binding pocket covered by a pH-sensitive āLidā
Background
The yellow fever mosquito, Aedes aegypti, is the primary vector for the viruses that cause yellow fever, mostly in tropical regions of Africa and in parts of South America, and human dengue, which infects 100 million people yearly in the tropics and subtropics. A better understanding of the structural biology of olfactory proteins may pave the way for the development of environmentally-friendly mosquito attractants and repellents, which may ultimately contribute to reduction of mosquito biting and disease transmission.
Methodology
Previously, we isolated and cloned a major, female-enriched odorant-binding protein (OBP) from the yellow fever mosquito, AaegOBP1, which was later inadvertently renamed AaegOBP39. We prepared recombinant samples of AaegOBP1 by using an expression system that allows proper formation of disulfide bridges and generates functional OBPs, which are indistinguishable from native OBPs. We crystallized AaegOBP1 and determined its three-dimensional structure at 1.85 Ć
resolution by molecular replacement based on the structure of the malaria mosquito OBP, AgamOBP1, the only mosquito OBP structure known to date.
Conclusion
The structure of AaegOBP1 (ā=āAaegOBP39) shares the common fold of insect OBPs with six Ī±-helices knitted by three disulfide bonds. A long molecule of polyethylene glycol (PEG) was built into the electron-density maps identified in a long tunnel formed by a crystallographic dimer of AaegOBP1. Circular dichroism analysis indicated that delipidated AaegOBP1 undergoes a pH-dependent conformational change, which may lead to release of odorant at low pH (as in the environment in the vicinity of odorant receptors). A C-terminal loop covers the binding cavity and this ālidā may be opened by disruption of an array of acid-labile hydrogen bonds thus explaining reduced or no binding affinity at low pH
Circulating Levels of Adipocyte and Epidermal Fatty AcidāBinding Proteins in Relation to Nephropathy Staging and Macrovascular Complications in Type 2 Diabetic Patients
OBJECTIVEāTo investigate the relationships of serum adipocyte fatty acidābinding protein (A-FABP) and epidermal fatty acidābinding protein (E-FABP) with renal dysfunction and macrovascular complications in type 2 diabetic patients
Trib2 suppresses tumor initiation in Notch-driven T-ALL
Trib2 is highly expressed in human T cell acute lymphoblastic leukemia (T-ALL) and is a direct transcriptional target of the oncogenic drivers Notch and TAL1. In human TAL1-driven T-ALL cell lines, Trib2 is proposed to function as an important survival factor, but there is limited information about the role of Trib2 in primary T-ALL. In this study, we investigated the role of Trib2 in the initiation and maintenance of Notch-dependent T-ALL. Trib2 had no effect on the growth and survival of murine T-ALL cell lines in vitro when expression was blocked by shRNAs. To test the function of Trib2 on leukemogenesis in vivo, we generated Trib2 knockout mice. Mice were born at the expected Mendelian frequencies without gross developmental anomalies. Adult mice did not develop pathology or shortened survival, and hematopoiesis, including T cell development, was unperturbed. Using a retroviral model of Notch-induced T-ALL, deletion of Trib2 unexpectedly decreased the latency and increased the penetrance of T-ALL development in vivo. Immunoblotting of primary murine T-ALL cells showed that the absence of Trib2 increased C/EBPĪ± expression, a known regulator of cell proliferation, and did not alter AKT or ERK phosphorylation. Although Trib2 was suggested to be highly expressed in T-ALL, transcriptomic analysis of two independent T-ALL cohorts showed that low Trib2 expression correlated with the TLX1-expressing cortical mature T-ALL subtype, whereas high Trib2 expression correlated with the LYL1-expressing early immature T-ALL subtype. These data indicate that Trib2 has a complex role in the pathogenesis of Notch-driven T-ALL, which may vary between different T-ALL subtypes
Quantum Computing and Quantum Simulation with Group-II Atoms
Recent experimental progress in controlling neutral group-II atoms for
optical clocks, and in the production of degenerate gases with group-II atoms
has given rise to novel opportunities to address challenges in quantum
computing and quantum simulation. In these systems, it is possible to encode
qubits in nuclear spin states, which are decoupled from the electronic state in
the S ground state and the long-lived P metastable state on the
clock transition. This leads to quantum computing scenarios where qubits are
stored in long lived nuclear spin states, while electronic states can be
accessed independently, for cooling of the atoms, as well as manipulation and
readout of the qubits. The high nuclear spin in some fermionic isotopes also
offers opportunities for the encoding of multiple qubits on a single atom, as
well as providing an opportunity for studying many-body physics in systems with
a high spin symmetry. Here we review recent experimental and theoretical
progress in these areas, and summarise the advantages and challenges for
quantum computing and quantum simulation with group-II atoms.Comment: 11 pages, 7 figures, review for special issue of "Quantum Information
Processing" on "Quantum Information with Neutral Particles
Charmless Exclusive Baryonic B Decays
We present a systematical study of two-body and three-body charmless baryonic
B decays. Branching ratios for two-body modes are in general very small,
typically less than , except that \B(B^-\to p \bar\Delta^{--})\sim
1\times 10^{-6}. In general, due to
the large coupling constant for . For three-body modes we
focus on octet baryon final states. The leading three-dominated modes are with a branching ratio of
order for and
for . The penguin-dominated decays with strangeness
in the meson, e.g., and , have appreciable rates and the mass
spectrum peaks at low mass. The penguin-dominated modes containing a strange
baryon, e.g., , have
branching ratios of order . In contrast, the decay
rate of is smaller. We explain why some of
charmless three-body final states in which baryon-antibaryon pair production is
accompanied by a meson have a larger rate than their two-body counterparts:
either the pole diagrams for the former have an anti-triplet bottom baryon
intermediate state, which has a large coupling to the meson and the
nucleon, or they are dominated by the factorizable external -emission
process.Comment: 46 pages and 3 figures, to appear in Phys. Rev. D. Major changes are:
(i) Calculations of two-body baryonic B decays involving a Delta resonance
are modified, and (ii) Penguin-dominated modes B-> Sigma+N(bar)+p are
discusse
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