Talin and vinculin play distinct roles in filopodial motility in the neuronal growth cone.
AuthorsSydor AM, Su AL, Wang FS, Xu A, Jay DG
JournalJ Cell Biol
PubMed ID8794861
'Filopodial motility is critical for many biological processes, particularly for axon guidance. This motility is based on altering the F-actin-based cytoskeleton, but the mechanisms of how this occurs and the actin-associated proteins that function in this process remain unclear. We investigated two of these proteins found in filopodia, talin and ... More
Chromophore-assisted laser inactivation of a repulsive axonal guidance molecule.
AuthorsMüller BK, Jay DG, Bonhoeffer F
JournalCurr Biol
PubMed ID8939610
'BACKGROUND: The axons of retinal ganglion neurons from a precise topographic map in the optic tectum in the midbrain, and the guidance of retinal axons by directional cues in the tectum is crucial in this process. Several in vitro systems have been developed in order to identify the molecular basis ... More
Photosensitization of Staphylococcus aureus with malachite green isothiocyanate: inactivation efficiency and spectroscopic analysis.
AuthorsGolding PS, King TA, Maddocks L, Drucker DB, Blinkhorn AS
JournalJ Photochem Photobiol B
PubMed ID10093920
'The potential of malachite green isothiocyanate as a photosensitizer for the inactivation of bacteria has been evaluated. Samples of Staphylococcus aureus are treated with the dye and exposed to continuous-wave red light from a filtered xenon lamp. Reduction in cell viability is seen to increase with radiation dose, whilst non-photosensitized ... More
Spatial specificity of chromophore assisted laser inactivation of protein function.
AuthorsLinden KG, Liao JC, Jay DG
JournalBiophys J
PubMed ID1581504
'Chromophore assisted laser inactivation (CALI) is a new technique that selectively inactivates proteins of interest to elucidate their in vivo functions. This method has application to a wide array of biological questions. An understanding of aspects of the mechanism of CALI is required for its judicious application. A critical concern ... More
Chromophore-assisted laser inactivation of cellular proteins.
AuthorsBeermann AE, Jay DG
JournalMethods Cell Biol
PubMed ID7707977
Nanoscale probing of adsorbed species by tip-enhanced Raman spectroscopy.
AuthorsPettinger B, Ren B, Picardi G, Schuster R, Ertl G
JournalPhys Rev Lett
PubMed ID15089490
Tip-enhanced Raman spectroscopy (TERS) is based on the optical excitation of localized surface plasmons in the tip-substrate cavity, which provides a large but local field enhancement near the tip apex. We report on TERS with smooth single crystalline surfaces as substrates. The adsorbates were CN- ions at Au(111) and malachite ... More
Near-infrared fluorescence lifetime assay for serum glucose based on allophycocyanin-labeled concanavalin A.
AuthorsMcCartney LJ, Pickup JC, Rolinski OJ, Birch DJ
JournalAnal Biochem
PubMed ID11355853
We describe an assay scheme for glucose based on fluorescence resonance energy transfer (FRET) between concanavalin A (con A), labeled with the near-infrared fluorescent protein allophycocyanin (APC) as donor, and dextran labeled with malachite green (MG) as acceptor. Glucose competitively displaces dextran-MG and leads to reduction in FRET, assessed by ... More
Selective destruction of protein function by chromophore-assisted laser inactivation.
AuthorsJay DG
JournalProc Natl Acad Sci U S A
PubMed ID3399501
Chromophore-assisted laser inactivation of protein function has been achieved. After a protein binds a specific ligand or antibody conjugated with malachite green (C.I. 42,000), it is selectively inactivated by laser irradiation at a wavelength of light absorbed by the dye but not significantly absorbed by cellular components. Ligand-bound proteins in ... More
Lifetime-based sensing of glucose using energy transfer with a long lifetime donor.
AuthorsTolosa L, Szmacinski H, Rao G, Lakowicz JR
JournalAnal Biochem
PubMed ID9234903
We describe an optical assay for glucose based on the luminescence decay time of a long lifetime metal-ligand complex. Concanavalin A was covalently labeled with Ruthenium metal-ligand complex (RuCon A) which served as the donor. The acceptor was malachite green which was covalently linked to insulin. The malachite green insulin ... More
Asymmetric retraction of growth cone filopodia following focal inactivation of calcineurin.
AuthorsChang HY, Takei K, Sydor AM, Born T, Rusnak F, Jay DG
JournalNature
PubMed ID7544441
The neuronal growth cone is thought to be the site of decision making in nerve growth and guidance. One likely mechanism of how the growth cone translates various extracellular cues into directed motility involves rises in intracellular calcium. A variety of physiological cues, such as adhesion molecules and neurotransmitters, increases ... More
Myosin 1c and myosin IIB serve opposing roles in lamellipodial dynamics of the neuronal growth cone.
AuthorsDiefenbach TJ, Latham VM, Yimlamai D, Liu CA, Herman IM, Jay DG
JournalJ Cell Biol
PubMed ID12356865
The myosin family of motor proteins is implicated in mediating actin-based growth cone motility, but the roles of many myosins remain unclear. We previously implicated myosin 1c (M1c; formerly myosin I beta) in the retention of lamellipodia (Wang et al., 1996). Here we address the role of myosin II (MII) ... More
Chromophore-assisted laser inactivation of proteins is mediated by the photogeneration of free radicals.
AuthorsLiao JC, Roider J, Jay DG
JournalProc Natl Acad Sci U S A
PubMed ID8146171
Chromophore-assisted laser inactivation (CALI) is a technique that selectively inactivates proteins of interest to elucidate their in vivo functions. This method has application to a wide array of biological questions and an understanding of its mechanism is required for its judicious application. We report here that CALI is not mediated ... More
Physicochemical studies of the d(G3T4G3)*d(G3A4G3).d(C3T4C3) triple helix.
AuthorsScaria PV, Will S, Levenson C, Shafer RH
JournalJ Biol Chem
PubMed ID7706270
We have targeted the d(G3A4G3).d(C3T4C3) duplex for triplex formation with d(G3T4G3) in the presence of MgCl2. The resulting triple helix, d(G3T4G3)*d(G3-A4G3).d(C3T4C3), is considerably weaker than the related triplex, d(G3A4G3)*d(G3A4G3).d(C3T4C3), and melts in a biphasic manner, with the third strand dissociating at temperatures about 20-30 degrees C below that of the ... More
Cytochrome P450 fluorometric substrates: identification of isoform-selective probes for rat CYP2D2 and human CYP3A4.
AuthorsStresser DM, Turner SD, Blanchard AP, Miller VP, Crespi CL
JournalDrug Metab Dispos
PubMed ID12065444
We have tested a panel of 29 cDNA-expressed rat and human enzymes with 9 fluorometric substrates to determine the P450 isoform selectivity in the catalysis of the substrates to fluorescent products. The substrates examined were dibenzyl fluorescein, 7-benzyloxyquinoline (BQ), 3-cyano-7-ethoxycoumarin, 3-cyano-7-methoxycoumarin, 7-methoxy-4-trifluoromethylcoumarin, 3-[2-(N,N-diethyl-N-methylamino)ethyl]-7-methoxy-4-methylcoumarin (AMMC), 3-[2-(N,N-diethyl-N-methylamino)ethyl]-7-methoxy-4-trifluoromethylcoumarin, 7-benzyloxyresorufin, and 7-benzyloxy-4-trifluoromethylcoumarin (BFC). For ... More