Biosynthetic transport from the trans-Golgi network (TGN) to the plasma membrane (PM) is mediated by secretory vesicles. We analyzed secretory vesicle transport in real time using a GFP-tagged secretory protein, hCgB-GFP, consisting of human chromogranin B (hCgB) and green fluorescent protein (GFP). The fusion protein was expressed transiently in Vero cells or in a stable clone after induction ...

The unusual membrane motor protein prestin is essential for mammalian hearing and for the survival of cochlear outer hair cells. While prestin has been demonstrated to be a homooligomer, by Western blot and FRET analyses, the stoichiometry of self association is unclear. Prestin, coupled to the enhanced green fluorescent protein, was synthesized and membrane targeted in human embryonic kidney c...

Green fluorescent protein fused to human chromogranin B or neuropeptide Y was expressed in PC12 cells and caused bright, punctate fluorescence. The fluorescent points colocalized with the endogenous secretory granule marker dopamine beta-hydroxylase. Stimulation of live PC12 cells with elevated [K+], or of permeabilized PC12 cells with Ca2+, led to Ca2+-dependent loss of fluorescence from neuri...

Green fluorescent protein, GFP, is a spontaneously fluorescent protein isolated from coelenterates, such as the Pacific jellyfish, Aequorea victoria, or from the sea pansy, Renilla reniformis. Its role is to transduce the blue chemiluminescence of aequorin, into green fluorescent light by energy transfer. The molecular cloning of GFP cDNA and the expression of GFP as a functional transgene has ...

Photoconvertible fluorescent proteins (FPs) are recent additions to the biologists' toolbox for understanding the living cell. Like green fluorescent protein (GFP), monomeric EosFP is bright green in color but is efficiently photoconverted into a red fluorescent form using a mild violet-blue excitation. Here, we report mEosFP-based probes that localize to the cytosol, plasma membrane invaginati...

Green fluorescent protein (GFP) and herpes simplex virus type-I thymidine kinase (TK) are commonly used markers in gene transfer studies. The latter gene has also proven to be an effective tool in cancer "suicide" gene therapy. To facilitate rapid and reliable selection of cells expressing TK, we constructed a plasmid expressing a TK-green fluorescent protein fusion gene (TK-GFP). In this fusio...

eosinophil autofluorescence. J. Leukoc. Biol. 51:172-175. 12.Nokubo, M., M. Ohta, K. Kitani and I. Z.Nagy. Identification of protein-bound riboflavin in rat hepatocyte plasma membrane as a source of autofluorescence. Biochim. Biophys. Acta 981:303-308. 13.Olson, K.R., J.R. McIntosh and J.B. Olmsted. 1995. Analysis of MAP4 function in living cells using green fluorescent protein (GFP) chimeras. ...

In the nearly two decades since the popularization of green fluorescent protein (GFP), fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compend...

G protein-coupled receptor kinases (GRKs) phosphorylate opioid receptors, which eventually results in receptor sequestration. With respect to kappa-opioid receptors, it is known that internalization occurs in a species-specific manner. That is, the agonist-occupied human kappa-receptors will sequester whereas murine receptors fail to do so. This investigation concentrates on the internalization...