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G-protein-coupled receptor (GPCR) activity gradients evoke important cell behavior but there is a dearth of methods to induce such asymmetric signaling in a cell. Here we achieved reversible, rapidly switchable patterns of spatiotemporally restricted GPCR activity in a single cell. We recruited properties of nonrhodopsin opsins--rapid deactivation, distinct(More)
There is a dearth of approaches to experimentally direct cell migration by continuously varying signal input to a single cell, evoking all possible migratory responses and quantitatively monitoring the cellular and molecular response dynamics. Here we used a visual blue opsin to recruit the endogenous G-protein network that mediates immune cell migration.(More)
G-protein βγ subunits translocate reversibly from the plasma membrane to internal membranes on receptor activation. Translocation rates differ depending on the γ subunit type. There is limited understanding of the role of the differential rates of Gβγ translocation in modulating signaling dynamics in a cell. Bifurcation analysis of the calcium oscillatory(More)
The protein S2 has been isolated from the 30S subunit of Escherichia coli A19 ribosomes [Littlechild, J., & Malcolm, A.L. (1978) Biochemistry 17, 3363-3369]. This salt-extracted protein is soluble and does not aggregate at salt concentrations of 0.3-0.4 M as used under reconstitution conditions. This differs from the S2 protein extracted by the acetic acid(More)
A complex mixture of 21 proteins from the 30S ribosomal subunit of Escherichia coli was fractionated on a cation-exchanger, then further separated on a C8 reversed-phase column. A set of 14 proteins were purified to homogeneity. The same protein mixture was also analysed on a C8 RPC column using a triethylamine phosphate (TEAP, pH2.2)/acetonitrile or a(More)
Proteins L1, L9, L25, and L30, purified by a nondenaturing method from the 50S ribosomal subunit of Escherichia coli A19, have been characterized. The four proteins were studied under conditions which resemble those used for reconstitution experiments. These proteins have S020,W values of 2.0 S, 1.8 S, 1.8 S, and 1.0 S and D20,W values of 8.4 X 10(-7), 9.0(More)
The shapes of proteins S3 and S17 purified from the 30 S subunit of Escherichia coli A19 were studied by hydrodynamic methods. The proteins have s020,w values of 2.1 +/- 0.1 S and 1.2 +/- 0.1 S and D020,w values of 7.4 +/- 0.5 . 10(-7) cm2/s and 11.4 +/- 0.6 . 10(-7) cm2/s. The respective molecular weights determined by sedimentation equilibrium are 25 800(More)