2,015 research outputs found
Nanomechanical and thermophoretic analyses of the nucleotide-dependent interactions between the AAA+ subunits of magnesium chelatase
In chlorophyll biosynthesis, the magnesium
chelatase enzyme complex catalyzes the insertion of a Mg2+
ion into protoporphyrin IX. Prior to this event, two of the three
subunits, the AAA+ proteins ChlI and ChlD, form a ChlID−
MgATP complex. We used microscale thermophoresis to
directly determine dissociation constants for the I-D subunits
from Synechocystis, and to show that the formation of a ChlID−
MgADP complex, mediated by the arginine finger and the
sensor II domain on ChlD, is necessary for the assembly of the
catalytically active ChlHID−MgATP complex. The N-terminal
AAA+ domain of ChlD is essential for complex formation, but
some stability is preserved in the absence of the C-terminal
integrin domain of ChlD, particularly if the intervening polyproline linker region is retained. Single molecule force spectroscopy
(SMFS) was used to determine the factors that stabilize formation of the ChlID−MgADP complex at the single molecule level;
ChlD was attached to an atomic force microscope (AFM) probe in two different orientations, and the ChlI subunits were
tethered to a silica surface; the probability of subunits interacting more than doubled in the presence of MgADP, and we show
that the N-terminal AAA+ domain of ChlD mediates this process, in agreement with the microscale thermophoresis data. Analysis
of the unbinding data revealed a most probable interaction force of around 109 pN for formation of single ChlID−MgADP
complexes. These experiments provide a quantitative basis for understanding the assembly and function of the Mg chelatase
complex
Protein sizing with Differential Dynamic Microscopy
Introduced more than fifty years ago, dynamic light scattering is routinely
used to determine the size distribution of colloidal suspensions, as well as of
macromolecules in solution, such as proteins, nucleic acids, and their
complexes. More recently, differential dynamic microscopy has been proposed as
a way to perform dynamic light scattering experiments with a microscope, with
much less stringent constraints in terms of cleanliness of the optical
surfaces, but a potentially lower sensitivity due to the use of camera-based
detectors. In this work, we push bright-field differential dynamic microscopy
beyond known limits and show it to be sufficiently sensitive to size small
macromolecules in diluted solutions. By considering solutions of three
different proteins (Bovine Serum Albumin, Lysozyme, and Pepsin), we accurately
determine the diffusion coefficient and hydrodynamic radius of both single
proteins and small protein aggregates down to concentrations of a few
milligrams per milliliter. In addition, we present preliminary results showing
unexplored potential for the determination of virial coefficients. Our results
are in excellent agreement with the ones obtained in parallel with a
state-of-the-art commercial dynamic light scattering setup, showing that
differential dynamic microscopy represents a valuable alternative for rapid,
label-free protein sizing with an optical microscope
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Biological predictors of chemotherapy-induced peripheral neuropathy (CIPN): MASCC neurological complications working group overview.
Chemotherapy-induced peripheral neuropathy (CIPN) is a common and debilitating condition associated with a number of chemotherapeutic agents. Drugs commonly implicated in the development of CIPN include platinum agents, taxanes, vinca alkaloids, bortezomib, and thalidomide analogues. As a drug response can vary between individuals, it is hypothesized that an individual's specific genetic variants could impact the regulation of genes involved in drug pharmacokinetics, ion channel functioning, neurotoxicity, and DNA repair, which in turn affect CIPN development and severity. Variations of other molecular markers may also affect the incidence and severity of CIPN. Hence, the objective of this review was to summarize the known biological (molecular and genomic) predictors of CIPN and discuss the means to facilitate progress in this field
Structural and functional consequences of removing the N-terminal domain from the magnesium chelatase ChlH subunit of Thermosynechococcus elongatus
Magnesium chelatase (MgCH) initiates chlorophyll biosynthesis by catalysing the ATP-dependent insertion of Mg2+ into protoporphyrin. This large enzyme complex comprises ChlH, I and D subunits, with I and D involved in ATP hydrolysis, and H the protein that handles the substrate and product. The 148 kDa ChlH subunit has a globular N-terminal domain attached by a narrow linker to a hollow cage-like structure. Following deletion of this ~18 kDa domain from the Thermosynechoccus elongatus ChlH, we used single particle reconstruction to show that the apo- and porphyrin-bound forms of the mutant subunit consist of a hollow globular protein with three connected lobes; superposition of the mutant and native ChlH structures shows that, despite the clear absence of the N-terminal ‘head’ region, the rest of the protein appears to be correctly folded. Analyses of dissociation constants shows that the ΔN159ChlH mutant retains the ability to bind protoporphyrin and the Gun4 enhancer protein, although the addition of I and D subunits yields an extremely impaired active enzyme complex. Addition of the Gun4 enhancer protein, which stimulates MgCH activity significantly especially at low Mg2+ concentrations, partially reactivates the ΔN159ChlH–I–D mutant enzyme complex, suggesting that the binding site or sites for Gun4 on H do not wholly depend on the N-terminal domain
Paediatric radiology seen from Africa. Part I: providing diagnostic imaging to a young population
Article approval pendingPaediatric radiology requires dedicated equipment, specific precautions related to ionising radiation, and specialist knowledge. Developing countries face difficulties in providing adequate imaging services for children. In many African countries, children represent an increasing proportion of the population, and additional challenges follow from extreme living conditions, poverty, lack of parental care, and exposure to tuberculosis, HIV, pneumonia, diarrhoea and violent trauma. Imaging plays a critical role in the treatment of these children, but is expensive and difficult to provide. The World Health Organisation initiatives, of which the World Health Imaging System for Radiography (WHIS-RAD) unit is one result, needs to expand into other areas such as the provision of maintenance servicing. New initiatives by groups such as Rotary and the World Health Imaging Alliance to install WHIS-RAD units in developing countries and provide digital solutions, need support. Paediatric radiologists are needed to offer their services for reporting, consultation and quality assurance for free by way of teleradiology. Societies for paediatric radiology are needed to focus on providing a volunteer teleradiology reporting group, information on child safety for basic imaging, guidelines for investigations specific to the disease spectrum, and solutions for optimising imaging in children
Trauma history and depression predict incomplete adherence to antiretroviral therapies in a low income country.
As antiretroviral therapy (ART) for HIV becomes increasingly available in low and middle income countries (LMICs), understanding reasons for lack of adherence is critical to stemming the tide of infections and improving health. Understanding the effect of psychosocial experiences and mental health symptomatology on ART adherence can help maximize the benefit of expanded ART programs by indicating types of services, which could be offered in combination with HIV care. The Coping with HIV/AIDS in Tanzania (CHAT) study is a longitudinal cohort study in the Kilimanjaro Region that included randomly selected HIV-infected (HIV+) participants from two local hospital-based HIV clinics and four free-standing voluntary HIV counselling and testing sites. Baseline data were collected in 2008 and 2009; this paper used data from 36 month follow-up interviews (N = 468). Regression analyses were used to predict factors associated with incomplete self-reported adherence to ART. INCOMPLETE ART ADHERENCE WAS SIGNIFICANTLY MORE LIKELY TO BE REPORTED AMONGST PARTICIPANTS WHO EXPERIENCED A GREATER NUMBER OF CHILDHOOD TRAUMATIC EVENTS: sexual abuse prior to puberty and the death in childhood of an immediate family member not from suicide or homicide were significantly more likely in the non-adherent group and other negative childhood events trended toward being more likely. Those with incomplete adherence had higher depressive symptom severity and post-traumatic stress disorder (PTSD). In multivariable analyses, childhood trauma, depression, and financial sacrifice remained associated with incomplete adherence.\ud
This is the first study to examine the effect of childhood trauma, depression and PTSD on HIV medication adherence in a low income country facing a significant burden of HIV. Allocating spending on HIV/AIDS toward integrating mental health services with HIV care is essential to the creation of systems that enhance medication adherence and maximize the potential of expanded antiretroviral access to improve health and reduce new infections
Measurement of Longitudinal Spin Transfer to Lambda Hyperons in Deep-Inelastic Lepton Scattering
Spin transfer in deep-inelastic Lambda electroproduction has been studied
with the HERMES detector using the 27.6 GeV polarized positron beam in the HERA
storage ring. For an average fractional energy transfer = 0.45, the
longitudinal spin transfer from the virtual photon to the Lambda has been
extracted. The spin transfer along the Lambda momentum direction is found to be
0.11 +/- 0.17 (stat) +/- 0.03 (sys); similar values are found for other
possible choices for the longitudinal spin direction of the Lambda. This result
is the most precise value obtained to date from deep-inelastic scattering with
charged lepton beams, and is sensitive to polarized up quark fragmentation to
hyperon states. The experimental result is found to be in general agreement
with various models of the Lambda spin content, and is consistent with the
assumption of helicity conservation in the fragmentation process.Comment: 8 pages, 3 figures; new version has an expanded discussion and small
format change
Nuclear transparency from quasielastic A(e,e'p) reactions uo to Q^2=8.1 (GeV/c)^2
The quasielastic (e,ep) reaction was studied on targets of
deuterium, carbon, and iron up to a value of momentum transfer of 8.1
(GeV/c). A nuclear transparency was determined by comparing the data to
calculations in the Plane-Wave Impulse Approximation. The dependence of the
nuclear transparency on and the mass number was investigated in a
search for the onset of the Color Transparency phenomenon. We find no evidence
for the onset of Color Transparency within our range of . A fit to the
world's nuclear transparency data reflects the energy dependence of the free
proton-nucleon cross section.Comment: 11 pages, 6 figure
Measurement of the Neutron Spin Structure Function with a Polarized ^3He Target
Results are reported from the HERMES experiment at HERA on a measurement of
the neutron spin structure function in deep inelastic scattering
using 27.5 GeV longitudinally polarized positrons incident on a polarized
He internal gas target. The data cover the kinematic range
and . The integral evaluated at a fixed of is . Assuming Regge behavior at low , the first
moment is .Comment: 4 pages TEX, text available at
http://www.krl.caltech.edu/preprints/OAP.htm
Observation of a Single-Spin Azimuthal Asymmetry in Semi-Inclusive Pion Electro-Production
Single-spin asymmetries for semi-inclusive pion production in deep-inelastic
scattering have been measured for the first time. A significant target-spin
asymmetry of the distribution in the azimuthal angle phi of the pion relative
to the lepton scattering plane was observed for pi+ electro-production on a
longitudinally polarized hydrogen target. The corresponding analyzing power in
the sin(phi) moment of the cross section is 0.022 +/- 0.005 +/- 0.003. This
result can be interpreted as the effect of terms in the cross section involving
chiral-odd spin distribution functions in combination with a time-reversal-odd
fragmentation function that is sensitive to the transverse polarization of the
fragmenting quark.Comment: 5 pages of RevTex, 3 ps figures, 2 table
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