Interaction of Haloarchaeal Gas Vesicle Proteins Determined by Split-GFP

Living synaptic vesicle marker: synaptotagmin-GFP.

Synapses are the site of chemical communication between neurons and between neurons and muscles. The synaptic vesicle (SV) is a prominent presynaptic organelle which contains chemical neurotransmitters and fuses with the plasma membrane to mediate neurotransmission. There are about 50 or so synaptic proteins which are either integral vesicle membrane proteins (e.g., synaptotagmin, syt; and syna...

A Split-GFP Gateway Cloning System for Topology Analyses of Membrane Proteins in Plants

To understand the function of membrane proteins, it is imperative to know their topology. For such studies, a split green fluorescent protein (GFP) method is useful. GFP is barrel-shaped, consisting of 11 β-sheets. When the first ten β-sheets (GFP1-10) and the 11th β-sheet (GFP11) are expressed from separate genes they will self-assembly and reconstitute a fluorescent GFP protein. However, this...

INTERACTION OF PROTEINS WITH SMALL MOLECULES

Screening libraries to identify proteins with desired binding activities using a split-GFP reassembly assay.

Designer protein modules, which bind specifically to a desired target, have numerous potential applications. One approach to creating such proteins is to construct and screen libraries. Here we present a detailed description of using a split-GFP reassembly assay to screen libraries and identify proteins with novel binding properties. Attractive features of the split-GFP based screen are the abs...

A New Protein-Protein Interaction Sensor Based on Tripartite Split-GFP Association

Monitoring protein-protein interactions in living cells is key to unraveling their roles in numerous cellular processes and various diseases. Previously described split-GFP based sensors suffer from poor folding and/or self-assembly background fluorescence. Here, we have engineered a micro-tagging system to monitor protein-protein interactions in vivo and in vitro. The assay is based on tripart...