A HIGHLY REACTIVE HETEROATOM ANALOG OF RETINAL AND ITS INTERACTION WITH BACTERIORHODOPSIN

Abstract— C₁₈ formate ester (5) [2-(6-methyl-8-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3E,5E,7E-octatrienyl formate], a highly reactive analog of retinal, was synthesized and its interaction with bacterioopsin studied. The formate ester, in the absence of purple or bleached membrane, undergoes very rapid reaction (t_l/2= 0.9 min) in neutral buffer but with membrane present it diffuses more rapidly into the membrane where it reacts slowly. Incorporation of 5 in the membrane results in a 38 nm (3900 cm^-1) red shift which remains after reconstitution with retinal. Similar experiments with the corresponding C₁₈ alcohol (4) results in a red shift, but this absorption blue shifts upon reconstitution with retinal. Washing the formate ester-treated membrane with bovine serum albumin or the corresponding lyophilized preparation with hexane, treatments that remove retinal oxime, fails to remove the UV-visible absorption, suggesting that a covalent bond between the C₁₈ moiety and a nucleophilic group of the protein has probably formed.     

STUDIES OF ADHESION OF METAL PARTICLES TO SILICA PARTICLES BASED ON ZETA POTENTIAL MEASUREMENTS

The zeta-potentials of silica, copper, platinum and gold particles have been measured as a function of pH. The isoelectric points were found to be at pH 3.0, 5.8, 3.0 and 3.5, respectively. In the pH range 3.0 to 5.8 copper and silica particles are oppositely charged and accordingly the coating of silica with copper particles could be demonstrated. In the case of gold and platinum the sign of the charge is such that direct adhesion to silica particles cannot be expected and this was also demonstrated in the case of platinum.     

Detection of NMR signals with a radio-frequency atomic magnetometer

We demonstrate detection of proton NMR signals with a radio-frequency (rf) atomic magnetometer tuned to the NMR frequency of 62 kHz. High-frequency operation of the atomic magnetometer makes it relatively insensitive to ambient magnetic field noise. We obtain magnetic field sensitivity of 7 fT/Hz1/2 using only a thin aluminum shield. We also derive an expression for the fundamental sensitivity limit of a surface inductive pick-up coil as a function of frequency and find that an atomic rf magnetometer is intrinsically more sensitive than a coil of comparable size for frequencies below about 50 MHz.     

Tunable atomic magnetometer for detection of radio-frequency magnetic fields

We describe an alkali-metal magnetometer for detection of weak magnetic fields in the radio-frequency (rf) range. High sensitivity is achieved by tuning the Zeeman resonance of alkali atoms to the rf frequency and partially suppressing spin-exchange collisions in the alkali-metal vapor. We demonstrate magnetic field sensitivity of 2 fT/Hz(1/2) at a frequency of 99 kHz with a resonance width of 400 Hz. We also derive a simple analytic expression for the fundamental limit on the sensitivity of the rf magnetometer and show that a sensitivity of about 0.01 fT/Hz(1/2) can be achieved in a practical system with a measurement volume of 200 cm3.     

13-Acetoxy-13-desmethylretinal-14-3H: Synthesis and Interaction with Bacterioopsin

The title compound (4) was synthesized in three steps from 1,1-dimethoxy-7-methyl-9-(2′,6′,6′-trimethyl-1-cy-clohexen-1-yl)-4E,6E,8E-nonatrien-3-one (1). Interaction of 4 with bacterioopsin produced a pigment absorbing at 573 nm, which, on long standing in the dark, moves to 406 nm, mirroring the behavior of nonradioactive 4 with bacterioopsin (S. Seltzer, J. Org. Chem. 60, 1189–1194, 1993). Compound 4 was designed to test for the involvement of a nucleophilic side chain, presumably asp-212, in catalyzing the dark cis-trans isomerization of bound retinal, through its attack at C-13. Such a mechanism is anticipated to lead to a cross-linked product where the nucleophile replaces the 13-acetoxy group when 4 is substituted for the natural chromophore. The protein, reconstituted with 4, after extensive washing, solubilization in dimethylsulfoxide and extensive dialysis retains its radioactivity suggesting the establishment of a cross-link. Solution in 6 M urea, however, results in substantial loss of radioactivity, suggesting that the unfolded labeled protein suffers hydrolysis of the very labile 13-enolester group followed by proton exchange.     

<Emphasis Type="Italic">N</Emphasis>-arachidonoyl glycine,an abundant endogenous lipid,potently drives directed cellular migration through GPR18, the putative abnormal cannabidiol receptor

Background  

Microglia provide continuous immune surveillance of the CNS and upon activation rapidly change phenotype to express receptors that respond to chemoattractants during CNS damage or infection. These activated microglia undergo directed migration towards affected tissue. Importantly, the molecular species of chemoattractant encountered determines if microglia respond with pro- or anti-inflammatory behaviour, yet the signaling molecules that trigger migration remain poorly understood. The endogenous cannabinoid system regulates microglial migration via CB₂ receptors and an as yet unidentified GPCR termed the 'abnormal cannabidiol' (Abn-CBD) receptor. Abn-CBD is a synthetic isomer of the phytocannabinoid cannabidiol (CBD) and is inactive at CB₁ or CB₂ receptors, but functions as a selective agonist at this G_i/o-coupled GPCR. N-arachidonoyl glycine (NAGly) is an endogenous metabolite of the endocannabinoid anandamide and acts as an efficacious agonist at GPR18. Here, we investigate the relationship between NAGly, Abn-CBD, the unidentified 'Abn-CBD' receptor, GPR18, and BV-2 microglial migration.