Active water in protein-protein communication within the membrane: the case of SRII-HtrII signal relay.

We detect internal water molecules in a membrane-embedded receptor-transducer complex and demonstrate water structure changes during formation of the signaling state. Time-resolved FTIR spectroscopy reveals stimulus-induced repositioning of one or more structurally active water molecules to a significantly more hydrophobic environment in the signaling state of the sensory rhodopsin II (SRII)-transducer (HtrII) complex. These waters, distinct from bound water molecules within the SRII receptor, appear to be in the middle of the transmembrane interface region near the Tyr199(SRII)-Asn74(HtrII) hydrogen bond. We conclude that water potentially plays an important role in the SRII --> HtrII signal transfer mechanism in the membrane's hydrophobic core.

Signal relay between two integral membrane proteins: The sensory rhodopsin I/HtrI transducer molecular complex

The molecular complex of sensory rhodopsin I (SRI) and its transducer HtrI mediate color-sensitive phototaxis in the archaeon Halobacterium salinarum. Orange light causes an attractant response by a one-photon reaction and white light causes a repellent response by a two-photon reaction. Three aspects of this molecular complex were explored: (i) We determined the stoichiometry of SRI and HtrI to be 2:2 by gene fusion analysis. A SRI-HtrI fusion protein was expressed in H. salinarum and shown to mediate 1-photon and 2-photon phototaxis responses comparable to wild-type complex. Disulfide crosslinking demonstrated that the fusion protein is a homodimer in the membrane. Measurement of photochemical reaction kinetics and pH titration of absorption spectra established that both SRI domains are complexed to HtrI in the fusion protein, and therefore the stoichiometry is 2:2. (ii) Cytoplasmic channel closure of SRI by HtrI, an important aspect of their interaction, was investigated by incremental HtrI truncation. We found that binding of the membrane-embedded portion of HtrI is insufficient for channel closure, whereas cytoplasmic extension of the second HtrI transmembrane helix by 13 residues blocks proton conduction through the channel as well as full-length HtrI. The closure activity is localized to 5 specific residues, each of which incrementally contributes to reduction of proton conductivity. Moreover, these same residues in the dark incrementally and proportionally increase the pKa of the Asp76 counterion to the protonated Schiff base chromophore. We conclude that this critical region of HtrI alters the dark conformation of SRI as well as light-induced channel opening. (iii) We developed a procedure for reconstituting HtrI-free SRI and the SRI/HtrI complex into liposomes, which exhibit photocycles with opened and closed cytoplasmic channels, respectively, as in the membrane. This opens the way for study of the light-induced conformational change and the interaction in vitro by fluorescence and spin-labeling. Single-cysteine mutations were introduced into helix F of SRI, labeled with a nitroxide spin probe and a fluorescence probe, reconstituted into proteoliposomes, and light-induced conformational changes detected in the complex. The probe signals can now be used as the readout of signaling to analyze mutants and the kinetics of signal relay. ^

Spatial sampling methods for improved communication for wireless relay robots

Having reliable wireless communication in a network of mobile robots is an ongoing challenge, especially when the mobile robots are given tasks in hostile or harmful environments such as radiation environments in scientific facilities, tunnels with large metallic components and complicated geometries as found at CERN. In this paper, we propose a decentralised method for improving the wireless network throughput by optimizing the wireless relay robot position to receive the best wireless signal strength using implicit spatial diversity concepts and gradient-search algorithms. We experimentally demonstrate the effectiveness of the proposed solutions with a KUKA Youbot omni-directional mobile robot. The performance of the algorithms is compared under various scenarios in an underground scientific facility at CERN.

Generalized synchronization in relay systems with instantaneous coupling

We demonstrate the existence of generalized synchronization in systems that act as mediators between two dynamical units that, in turn, show complete synchronization with each other. These are the so-called relay systems. Specifically, we analyze the Lyapunov spectrum of the full system to elucidate when complete and generalized synchronization appear. We show that once a critical coupling strength is achieved, complete synchronization emerges between the systems to be synchronized, and at the same point, generalized synchronization with the relay system also arises. Next, we use two nonlinear measures based on the distance between phase-space neighbors to quantify the generalized synchronization in discretized time series. Finally, we experimentally show the robustness of the phenomenon and of the theoretical tools here proposed to characterize it.

A Novel Ras-interacting Protein Required for Chemotaxis and Cyclic Adenosine Monophosphate Signal Relay in Dictyostelium

We have identified a novel Ras-interacting protein from Dictyostelium, RIP3, whose function is required for both chemotaxis and the synthesis and relay of the cyclic AMP (cAMP) chemoattractant signal. rip3 null cells are unable to aggregate and lack receptor activation of adenylyl cyclase but are able, in response to cAMP, to induce aggregation-stage, postaggregative, and cell-type-specific gene expression in suspension culture. In addition, rip3 null cells are unable to properly polarize in a cAMP gradient and chemotaxis is highly impaired. We demonstrate that cAMP stimulation of guanylyl cyclase, which is required for chemotaxis, is reduced ∼60% in rip3 null cells. This reduced activation of guanylyl cyclase may account, in part, for the defect in chemotaxis. When cells are pulsed with cAMP for 5 h to mimic the endogenous cAMP oscillations that occur in wild-type strains, the cells will form aggregates, most of which, however, arrest at the mound stage. Unlike the response seen in wild-type strains, the rip3 null cell aggregates that form under these experimental conditions are very small, which is probably due to the rip3 null cell chemotaxis defect. Many of the phenotypes of the rip3 null cell, including the inability to activate adenylyl cyclase in response to cAMP and defects in chemotaxis, are very similar to those of strains carrying a disruption of the gene encoding the putative Ras exchange factor AleA. We demonstrate that aleA null cells also exhibit a defect in cAMP-mediated activation of guanylyl cyclase similar to that of rip3 null cells. A double-knockout mutant (rip3/aleA null cells) exhibits a further reduction in receptor activation of guanylyl cyclase, and these cells display almost no cell polarization or movement in cAMP gradients. As RIP3 preferentially interacts with an activated form of the Dictyostelium Ras protein RasG, which itself is important for cell movement, we propose that RIP3 and AleA are components of a Ras-regulated pathway involved in integrating chemotaxis and signal relay pathways that are essential for aggregation.

UM Men's Swimming, 1971 200 Medley Relay Team

[L-R Bob Zann, Tim Norlen, Larry Day, Ray McCullough]

Sprint Medley Relay Team 1979, UM Men's Track, Set world record of 3:22 at Michigan Relays in Lansing, braking record set by U-M team in 1976

{L-R: Charles Crouther, Andrew Bruce, Ken Gardner, Tim Thomas]

Sprint Medley Relay Team 1979, UM Men's Track, Set world record of 3:22 at Michigan Relays in Lansing, braking record set by U-M team in 1976

[L-R: Charles Crouther, Andrew Bruce, Ken Gardner, Tim Thomas]

Potential means of cost reduction in grade crossings motorist-warning control equipment. Volume II: comparison of solid state and relay devices and techniques. Final report.

Transportation Systems Center, Cambridge, Mass.

Potential means of cost reduction in grade crossings motorist-warning control equipment. Volume I: overview, technology survey and relay alternatives. Final report.

Transportation Systems Center, Cambridge, Mass.