The Saccharomyces cerevisiae PCD1 gene encodes a peroxisomal nudix hydrolase active toward coenzyme A and its derivatives.
AuthorsCartwright JL, Gasmi L, Spiller DG, McLennan AG
JournalJ Biol Chem
PubMed ID10922370
'The PCD1 nudix hydrolase gene of Saccharomyces cerevisiae has been cloned and the Pcd1p protein characterized as a diphosphatase (pyrophosphatase) with specificity for coenzyme A and CoA derivatives. Oxidized CoA disulfide is preferred over CoA as a substrate with K(m) and k(cat) values of 24 micrometer and 5.0 s(-1), respectively, ... More
Impaired integrin-mediated adhesion and signaling in fibroblasts expressing a dominant-negative mutant PTP1B.
AuthorsArregui CO, Balsamo J, Lilien J
JournalJ Cell Biol
PubMed ID9813103
'To investigate the role of nonreceptor protein tyrosine phosphatase 1B (PTP1B) in beta1-integrin- mediated adhesion and signaling, we transfected mouse L cells with normal and catalytically inactive forms of the phosphatase. Parental cells and cells expressing the wild-type or mutant PTP1B were assayed for (a) adhesion, (b) spreading, (c) presence ... More
Mitochondrial membrane potential is dependent on the oligomeric state of F1F0-ATP synthase supracomplexes.
AuthorsBornhövd C, Vogel F, Neupert W, Reichert AS
JournalJ Biol Chem
PubMed ID16551625
'The F1F0-ATP synthase in mitochondria, in addition to its function in energy transduction, has a structural role in determining cristae morphology. This depends on its ability to form dimeric and higher oligomeric supracomplexes. Here we show that mutants of the dimer-specific subunits e and g, which destabilize dimeric and oligomeric ... More
Role of MMM1 in maintaining mitochondrial morphology in Neurospora crassa.
AuthorsProkisch H, Neupert W, Westermann B
JournalMol Biol Cell
PubMed ID10982393
'Mmm1p is a protein required for maintenance of mitochondrial morphology in budding yeast. It was proposed that it is required to mediate the interaction of the mitochondrial outer membrane with the actin cytoskeleton. We report the cloning and characterization of MMM1 of the filamentous fungus Neurospora crassa, an organism that ... More
Intracellular localization of the radiation enhancer motexafin gadolinium using interferometric Fourier fluorescence microscopy.
AuthorsWoodburn KW
JournalJ Pharmacol Exp Ther
PubMed ID11356908
'Motexafin gadolinium (MGd) is a unique therapeutic agent that localizes in cancer cells and increases tumor response to ionizing radiation and certain chemotherapeutics. The in vitro intracellular localization, accumulation, and retention of MGd in murine EMT6 mammary sarcoma and Rif-1 fibrosarcoma cell lines were studied using interferometric Fourier fluorescence microscopy. ... More
Diffusion across the nuclear envelope inhibited by depletion of the nuclear Ca2+ store.
AuthorsStehno-Bittel L, Perez-Terzic C, Clapham DE
JournalScience
PubMed ID8525380
Intact, isolated nuclei and a nuclear membrane (ghost) preparation were used to study regulation of the movement of small molecules across the Xenopus laevis oocyte nuclear membrane. In contrast to models of the nuclear pore complex, which assume passive bidirectional diffusion of molecules less than 70 kilodaltons, diffusion of intermediate-sized ... More
The missing link in coronavirus assembly. Retention of the avian coronavirus infectious bronchitis virus envelope protein in the pre-Golgi compartments and physical interaction between the envelope and membrane proteins.
AuthorsLim KP, Liu DX
JournalJ Biol Chem
PubMed ID11278557
One missing link in the coronavirus assembly is the physical interaction between two crucial structural proteins, the membrane (M) and envelope (E) proteins. In this study, we demonstrate that the coronavirus infectious bronchitis virus E can physically interact, via a putative peripheral domain, with M. Deletion of this domain resulted ... More
Characterization of endoplasmic reticulum by co-localization of BiP and dicarbocyanine dyes.
AuthorsTerasaki M, Reese TS
JournalJ Cell Sci
PubMed ID1378452
The original concept of endoplasmic reticulum derived from the observation of a reticular network in cultured fibroblasts by electron microscopy of whole cells. It was previously reported that the fluorescent dye, DiOC6(3), stains a similar network as well as mitochondria and other organelles in living cells. Here, we investigate the ... More
Functional studies of the kidney of living animals using multicolor two-photon microscopy.
AuthorsDunn KW, Sandoval RM, Kelly KJ, Dagher PC, Tanner GA, Atkinson SJ, Bacallao RL, Molitoris BA
JournalAm J Physiol Cell Physiol
PubMed ID12176747
Optical microscopy, when applied to living animals, provides a powerful means of studying cell biology in the most physiologically relevant setting. The ability of two-photon microscopy to collect optical sections deep into biological tissues has opened up the field of intravital microscopy to high-resolution studies of the brain, lens, skin, ... More
Rhodamine B, a fluorescent probe for acidic organelles in denervated skeletal muscle.
AuthorsVult von Steyern F, Josefsson JO, Tågerud S
JournalJ Histochem Cytochem
PubMed ID8648087
We describe a very efficient method for fluorescent labeling of acidic structures in denervated skeletal muscle with rhodamine B. Rhodamine B at 50 ng/ml gave selective and distinct segmental labeling of denervated muscle fibers after 5-min incubation at room temperature. Labeling was also achieved at 4 degrees C. The labeling ... More
Integral membrane proteins of the nuclear envelope are dispersed throughout the endoplasmic reticulum during mitosis.
AuthorsYang L, Guan T, Gerace L
JournalJ Cell Biol
PubMed ID9182656
We have analyzed the fate of several integral membrane proteins of the nuclear envelope during mitosis in cultured mammalian cells to determine whether nuclear membrane proteins are present in a vesicle population distinct from bulk ER membranes after mitotic nuclear envelope disassembly or are dispersed throughout the ER. Using immunofluorescence ... More
Visualization of oleic acid-induced transdermal diffusion pathways using two-photon fluorescence microscopy.
AuthorsYu B, Kim KH, So PT, Blankschtein D, Langer R
JournalJ Invest Dermatol
PubMed ID12603859
In a novel application of dual-channel high-speed two-photon fluorescence microscopy, the skin autofluores-cence and the transdermal fluorescent model drug spatial distributions were imaged simultaneously over precisely the same spatial coordinates. The dual channels enable the detection of the fluorescence emission wavelengths characteristic of the endogenous (intrinsic) skin fluorophores, as well ... More
InsP3 signaling induces pulse-modulated Ca2+ signals in the nucleus of airway epithelial ciliated cells.
AuthorsQuesada I, Verdugo P
JournalBiophys J
PubMed ID15792976
The phenomenology of nuclear Ca(2+) dynamics has experienced important progress revealing the broad range of cellular processes that it regulates. Although several agonists can mobilize Ca(2+) from storage in the nuclear envelope (NE) to the intranuclear compartment (INC), the mechanisms of Ca(2+) signaling in the nucleus still remain uncertain. Here ... More
Subcellular distribution of normal and mutant vitamin D receptors in living cells. Studies with a novel fluorescent ligand.
AuthorsBarsony J, Renyi I, McKoy W
JournalJ Biol Chem
PubMed ID9038191
To understand the subcellular localization of the vitamin D receptor (VDR) and to measure VDR content in single cells, we recently developed a fluorescent labeled ligand, 4,4-difluoro-4-bora-3a, 4a-diaza-s-indacene (BODIPY)-calcitriol. This tagged hormone has intact biological activity, high affinity and specific binding to the receptor, and enhanced fluorescent emission upon receptor ... More
Evaluation of fluorescent probe surface intensities as an indicator of transdermal permeant distributions using wide-area two-photon fluorescence microscopy.
AuthorsYu B, Kim KH, So PT, Blankschtein D, Langer R
JournalJ Pharm Sci
PubMed ID14603481
The application of high-speed two-photon fluorescence microscopy (HTPM) to examine transdermal transport processes has enabled the noninvasive visualization of permeant spatial distributions over a larger, more clinically relevant wide area of the skin. Earlier studies demonstrated that the transdermal fluorescent probe distribution over a 2 x 2 mm skin area ... More
Molecular characterization of FMN1, the structural gene for the monofunctional flavokinase of Saccharomyces cerevisiae.
AuthorsSantos MA, Jimenez A, Revuelta JL
JournalJ Biol Chem
PubMed ID10887197
Flavokinase catalyzes the transfer of the gamma-phosphoryl group of ATP to riboflavin to form the flavocoenzyme FMN. Consistent patterns of sequence similarities have identified the open reading frame of unknown function YDR236c as a candidate to encode flavokinase in Saccharomyces cerevisiae. In order to determine whether the product of this ... More
Shuttling of CTP:Phosphocholine cytidylyltransferase between the nucleus and endoplasmic reticulum accompanies the wave of phosphatidylcholine synthesis during the G(0) --> G(1) transition.
The transition from quiescence (G(0)) into the cell division cycle is marked by accelerated phospholipid turnover. We examined the rates of phosphatidylcholine (PC) synthesis and the activity, membrane affinity, and intracellular localization of the rate-limiting enzyme in the synthesis of PC, CTP:phosphocholine cytidylyltransferase (CT) during this transition. The addition of ... More