tRNA-linked molecular beacons for imaging mRNAs in the cytoplasm of living cells.
AuthorsMhlanga MM, Vargas DY, Fung CW, Kramer FR, Tyagi S
JournalNucleic Acids Res
PubMed ID15809226
'When oligonucleotide probes are microinjected into cells to image the distribution of RNAs, they are rapidly sequestered into the nucleus. As a result, it is difficult to detect mRNAs in the cytoplasm of living cells. We were able to overcome this process by attaching tRNA transcripts to the probes. We ... More
Wavelength-shifting molecular beacons.
AuthorsTyagi S, Marras SA, Kramer FR
JournalNat Biotechnol
PubMed ID11062440
'We describe wavelength-shifting molecular beacons, which are nucleic acid hybridization probes that fluoresce in a variety of different colors, yet are excited by a common monochromatic light source. The twin functions of absorption of energy from the excitation light and emission of that energy in the form of fluorescent light ... More
Purification and fluorescent labeling of the human serotonin transporter.
AuthorsRasmussen SG, Gether U
JournalBiochemistry
PubMed ID15736959
'To establish a purification procedure for the human serotonin transporter (hSERT) we expressed in Sf9 insect cells an epitope-tagged version of the transporter containing a FLAG epitope at the N-terminus and a polyhistidine tail at the C-terminus (FLAG-hSERT-12H). For purification, the transporter was solubilized in digitonin followed by nickel affinity ... More
Electrostatic sequestration of PIP2 on phospholipid membranes by basic/aromatic regions of proteins.
AuthorsGambhir A, Hangyás-Mihályné G, Zaitseva I, Cafiso DS, Wang J, Murray D, Pentyala SN, Smith SO, McLaughlin S
JournalBiophys J
PubMed ID15041659
The basic effector domain of myristoylated alanine-rich C kinase substrate (MARCKS), a major protein kinase C substrate, binds electrostatically to acidic lipids on the inner leaflet of the plasma membrane; interaction with Ca2+/calmodulin or protein kinase C phosphorylation reverses this binding. Our working hypothesis is that the effector domain of ... More