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Two different types of gap junction proteins, beta 1- and beta 2-connexin, were expressed in insect cells, either singly or together, using infection with recombinant baculovirus. Membrane fractions enriched in gap junction proteins were isolated, and connexons (hemichannels) were solubilized with detergent. These solubilized connexons were then run out on(More)
During normal development, rainbow trout undergo a shift in red muscle contraction kinetics and swimming kinematics. Young trout parr have faster muscle kinetics and faster tailbeat frequency during swimming than older, larger juvenile trout. In this study, the thyroid hormone thyroxine (T(4)) was used to induce these changes in trout parr. This allowed a(More)
BACKGROUND Voltage-gated K+ channels play an important role in the control of neuronal excitability and synaptic plasticity. Their low abundance and extraordinary heterogeneity have rendered their purification from natural sources difficult. We have previously cloned a voltage-gated K(+)-channel gene, Shaker, from Drosophila. The Shaker K(+)-channel(More)
The beta 2 gap junction protein (Cx26) was expressed in an insect cell line by infection with a baculovirus vector containing the rat beta 2 cDNA. Isolated beta 2 gap junction connexons were reconstituted into planar lipid bilayers. Single channel activity was observed with a unitary conductance of 35-45 pS in 200 mM KCl. Channels with conductance values of(More)
A recombinant baculovirus containing a mouse 5-hydroxytryptamine3 (5-HT3) receptor subunit cDNA under the control of the polyhedrin promoter was shown to direct the production of large amounts of functional 5-HT3 receptor in insect cells, as assayed by Western blotting and ligand binding. After solubilization, the receptor was purified to homogeneity by(More)
This paper reports methods we have developed to solubilize gap junction channels, or connexons, from isolated gap junctions and to purify them in milligram quantities. Two sources of material are used: rat liver gap junctions and gap junctions produced by infecting insect cells with a baculovirus containing the cDNA for human liver beta 1 protein (connexin(More)
Gap junctions are regions of contact between adjacent cells, consisting of arrays of channels linking the cell interiors. The channels are formed by polypeptides called connexins; the amino acid sequences of many different connexins are known, and they are thought to resemble each other closely in tertiary and quarternary structure. Single channels have(More)
Juvenile stages of rainbow trout, smaller parr and older juveniles, termed smolts, show differences in red muscle contractile properties: parr red muscle has faster kinetics and a faster maximum shortening velocity than smolt red muscle. A developmental reduction in the number of MHC isoforms as detected by SDS-PAGE between parr and smolt has also been(More)
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