A5039383940- Microtubule and mitosis dynamics
- Cellular transport and secretion
- Cellular Mechanics and Interactions
- Hemoglobin structure and function
- Neuropeptides and Animal Physiology
- Hearing, Cochlea, Tinnitus, Genetics
- Lipid Membrane Structure and Behavior
- Micro and Nano Robotics
- Nanopore and Nanochannel Transport Studies
- Electrochemical sensors and biosensors
- Protein purification and stability
- Receptor Mechanisms and Signaling
- Cell Adhesion Molecules Research
- Photosynthetic Processes and Mechanisms
- Heme Oxygenase-1 and Carbon Monoxide
- Ear Surgery and Otitis Media
- Peptidase Inhibition and Analysis
- Spectroscopy Techniques in Biomedical and Chemical Research
- Tribology and Wear Analysis
- Advanced Electron Microscopy Techniques and Applications
- Field-Flow Fractionation Techniques
- Enzyme function and inhibition
- Hearing Loss and Rehabilitation
- Silicone and Siloxane Chemistry
- Trace Elements in Health
National Institutes of Health
2012-2024
National Institute on Deafness and Other Communication Disorders
2022-2024
Mayo Clinic in Florida
2020-2023
WinnMed
2020-2023
Mayo Clinic in Arizona
2019-2020
National Institute of Neurological Disorders and Stroke
2012-2016
Interuniversity Consortium for Magnetic Resonance
2009
University of Florence
2009
National Institutes of Natural Sciences
2004-2007
Okazaki Institute for Integrative Bioscience
2004-2007
Kinesin-5 motors organize mitotic spindles by sliding apart microtubules. They are homotetramers with dimeric motor and tail domains at both ends of a bipolar minifilament. Here, we describe regulatory mechanism involving direct binding between its fundamental role in microtubule sliding. tails decrease microtubule-stimulated ATP-hydrolysis specifically engaging the nucleotide-free or ADP states. Cryo-EM reveals that stabilizes an open domain ATP-active site. Full-length undergo slow...
G protein-coupled receptor kinases (GRKs) play an important role in the desensitization of protein-mediated signaling receptors (GPCRs). The level interest mapping their phosphorylation sites has increased because recent studies suggest that differential pattern distinct biological consequences. In vitro experiments using well-controlled systems are useful for deciphering complexity these physiological reactions and understanding targeted event. Here, we report on class A GPCR neurotensin 1...
Hereditary deafness and retinal dystrophy are each genetically heterogenous clinically variable. Three small unrelated families segregating the combination of were studied by exome sequencing (ES). The proband Family 1 was found to be compound heterozygous for NM_004525.3: LRP2: c.5005A > G, p.(Asn1669Asp) c.149C p.(Thr50Ser). In 2, two sisters LRP2 variants, p.(Tyr3933Cys) an experimentally confirmed c.7715 + 3A T consensus splice-altering variant. 3, is a donor splice site variant...
We report the solution NMR structures of N-domain Menkes protein (ATP7A) in ATP-free and ATP-bound forms. The consist a twisted antiparallel six-stranded beta-sheet flanked by two pairs alpha-helices. A loop 50 amino acids located between beta 3 4 is disordered mobile on subnanosecond time scale. ATP binds with an affinity constant (1.2 +/- 0.1) x 10(4) m(-1) exchanges rate order 1 10(3) s(-1). ATP-binding cavity considerably affected presence ligand, resulting more compact conformation than...
CooA is a CO-sensing transcriptional activator that contains b-type heme as the active site for sensing its physiological effector, CO. In this study, spectroscopic and redox properties of new homologue from Carboxydothermus hydrogenoformans (Ch-CooA) were studied. Spectroscopic mutagenesis studies revealed His-82 N-terminal α-amino group axial ligands Fe(III) Fe(II) hemes in Ch-CooA was replaced by CO upon binding. Two neutral ligands, group, are coordinated to Ch-CooA, whereas two...
Kinesin-5 crosslinks and slides apart microtubules to assemble, elongate, maintain the mitotic spindle. is a tetramer, where two N-terminal motor domains are positioned at each end of motor, coiled-coil stalk organized into tetrameric bundle through bipolar assembly (BASS) domain. To dissect function individual structural elements we constructed minimal kinesin-5 tetramer (mini-tetramer). We determined x-ray structure extended, 34-nm BASS Guided by these studies, generated active...
The UV and visible resonance Raman spectra are reported for CooA from Rhodospirillum rubrum, which is a transcriptional regulator activated by growth in CO atmosphere. binding to heme its sensor domain causes rearrangement of DNA-binding domain, allowing DNA with specific sequence. domains linked hinge region that follows long C-helix. bands arising Trp-110 the C-helix revealed local movement around upon binding. indole side chain Trp-110, exposed solvent CO-free ferrous state, becomes...
Abstract Due to their unique properties, tunable size, and ability provide a near native lipid environment, nanodiscs have found widespread use for the structural functional studies of reconstituted membrane proteins. They also been developed, albeit in few applications, therapeutic biomedical use. For these it is essential characterize nanodisc preparations terms monodispersity, composition, as can influence properties protein interest. Of many biophysical methods utilized study...
Hearing depends on intricate morphologies and mechanical properties of diverse inner ear cell types. The individual contributions various types into the organ Corti mechanisms their integration are yet largely unknown. Using sub-100-nm spatial resolution atomic force microscopy (AFM), we mapped Young’s modulus (stiffness) apical surface different cells freshly dissected P5–P6 cochlear epithelium from wild-type mice lacking either Trio F-actin binding protein (TRIOBP) isoforms 4 5 or isoform...
CooA, a homodimeric haem-containing protein, is responsible for transcriptional regulation in response to carbon monoxide (CO). It has b-type haem as CO sensor. Upon binding the haem, CooA binds promoter DNA and activates expression of genes metabolism. from Carboxydothermus hydrogenoformans been overexpressed Escherichia coli, purified crystallized by vapour-diffusion method. The crystal belongs space group P2(1), with unit-cell parameters = 61.8, b 94.7, c 92.8 angstroms, beta 104.8...
Deafness in vertebrates is associated with variants of hundreds genes. Yet, many mutant genes causing rare forms deafness remain to be discovered. A consanguineous Pakistani family segregating nonsyndromic two sibships were studied using microarrays and exome sequencing. 1.2 Mb locus (DFNB128) on chromosome 5q11.2 encompassing six was identified. In one the this family, a novel homozygous recessive variant NM_005921.2:c.4460G>A p.(Arg1487His) kinase domain
Mutations of human TBC1D24 are associated with deafness, epilepsy, or DOORS syndrome (deafness, onychodystrophy, osteodystrophy, cognitive disability, and seizures). The causal relationships between variants the different clinical phenotypes not understood. Our hypothesis is that phenotypic heterogeneity missense mutations results, in part, from perturbed binding protein partners. To discover novel partners TBC1D24, we conducted yeast two-hybrid (Y2H) screen using mouse full-length as bait....
Abstract The conserved kinesin-5 bipolar tetrameric motors slide apart microtubules during mitotic spindle assembly and elongation. Kinesin-5 organization originates from its helical minifilament, which position the C-terminal tail domains of two subunits near N-terminal motor anti-parallel (Scholey et al, 2014). This unique structure enables to simultaneously engage transmit forces between them, for multiple organize via interactions microtubule sliding (Bodrug 2020). Here, we show how...
<title>Abstract</title> Deafness in vertebrates is associated with variants of hundreds genes. Yet, many genes causing rare forms deafness remain to be discovered. Microarrays and exome sequencing were used study a consanguineous Pakistani family segregating nonsyndromic two sibships. A 1.2 Mb locus (<italic>DFNB110</italic>) on chromosome 5q11.2 encompassing six was identified. In one the sibships this family, novel homozygous recessive variant NM_005921.2:c.4460G>A p.(Arg1487His) kinase...
Abstract Kinesin-5 motors organize mitotic spindles by sliding apart anti-parallel microtubules. They are homotetramers composed of two antiparallel dimers placing orthogonal motor and tail domains at opposite ends a bipolar minifilament. Here, we describe regulatory mechanism, involving direct binding the to domain reveal its fundamental role in microtubule motility. Biochemical analyses that down-regulates microtubule-activated ATP hydrolysis specifically engaging nucleotide-free or...