New spin labeling strategies have immense potential in studying protein structure and dynamics under physiological conditions with electron paramagnetic resonance (EPR) spectroscopy. Here, a new spin-labeled chemical recognition unit for switchable and concomitantly high affinity binding to His-tagged proteins was synthesized. In combination with an orthogonal site-directed spin label, this novel spin probe, Proxyl-trisNTA (P-trisNTA) allows the extraction of structural constraints within proteins and macromolecular complexes by EPR. By using the multisubunit maltose import system of E. coli: 1) the topology of the substrate-binding protein, 2) its substrate-dependent conformational change, and 3) the formation of the membrane multiprotein complex can be extracted. Notably, the same distance information was retrieved both in vitro and in situ allowing for site-specific spin labeling in cell lysates under in-cell conditions. This approach will open new avenues towards in-cell EPR.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/chem.201301921 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Reversible Perfusion Changes during Acute Attacks in Glucose Transporter Type 1 Deficiency Syndrome: A Pediatric Case Series.
AJNR Am J Neuroradiol
January 2025
Department of Pediatric Radiology and Neuroradiology (C.D., F.A., C.P., A.R.), Children's Hospital V. Buzzi, Milan, Italy.
Glucose transporter type 1 deficiency syndrome (GLUT1-DS) is an uncommon condition represented by an infantile-onset disorder, frequently arising from heterozygous mutations in the gene. Individuals with GLUT1-DS may present with early-onset seizures (typically manifesting before 4 years of age), developmental delay, and complex movement disorders. In fewer cases, stroke-like events or hemiplegic migraine-like symptoms are also reported, defined by unilateral paresis affecting 1 side of the body and/or one-half of the face, occasionally accompanied by speech impairment.
View Article and Find Full Text PDFBackground: Phase four of the Alzheimer's Disease Neuroimaging Initiative (ADNI4) began in 2023. This time-period corresponded to MRI vendors introducing product sequences with compressed sensing (CS), cross-vendor adoption of arterial spin-labelling (ASL) and multi-band slice excitation, and hardware improvements (head-coils, increased gradient amplitudes). These advances enabled the acquisition of new imaging measures and reduced scan times.
View Article and Find Full Text PDFBackground: Apolipoprotein ε4 allele (APOE4) is the strongest genetic risk factor for late-onset Alzheimer's disease (AD) with females having higher risk than males. Compared with non-carriers, cognitively normal, middle-aged APOE4 carriers have lower cerebral blood flow (CBF) decades before clinical symptoms appear. Early intervention to protect CBF would be critical for APOE4 carriers to mitigate AD progression.
View Article and Find Full Text PDFBackground: Non-invasive biofluid and MRI measures of blood-brain-barrier (BBB) dysfunction may aid early detection of cerebral small vessel disease (cSVD). Plasma markers of astrocytic function and injury, such as S100 calcium-binding protein B (S100b), have gained increased attention in relation to BBB integrity and cognition. Here we explored the inter-relationships between plasma S100b levels, an MRI measure of water exchange rate across the BBB (kw), and cognitive performance among older adults.
View Article and Find Full Text PDFBiomarkers.
Alzheimers Dement
December 2024
University of Kansas Medical Center, Kansas City, KS, USA.
Background: Cerebral blood flow (CBF) and glucose utilization have both proven sensitive biomarkers of brain function in Alzheimer's disease. However, while blood flow supplies glucose to cells to meet local demand, and therefore, are inter-related, the two aspects are physiologically distinct. Our goal was to conduct a region-to-region correlation of magnetic resonance imaging (MRI) and F-fluorodeoxyglucose positron emission tomography (FDG-PET) biomarkers of cerebral blood flow and glucose utilization to determine whether these physiologically distinct biomarkers yield functionally distinct information.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!