Role for bound water and CH-pi aromatic interactions in photosynthetic electron transfer

Photosystem I (PSI) is one of two photosynthetic reaction centers present in plants, algae, and cyanobacteria and catalyzes the reduction of ferredoxin and the oxidation of cytochrome c or plastocyanin. The PSI primary chlorophyll donor, which is oxidized in the primary electron-transfer events, is a heterodimer of chl a and a' called P700. It has been suggested that protein relaxation accompanies light-induced electron transfer in this reaction center (Dashdorj, N.; Xu, W.; Martinsson, P.; Chitnis, P. R.; Savikhin, S. Biophys. J. 2004, 86, 3121. Kim, S.; Sacksteder, C. A.; Bixby, K. A.; Barry, B. A. Biochemistry 2001, 40, 15384). To investigate the details of electron transfer and relaxation events in PSI, we have employed several experimental approaches. First, we report a pH-dependent viscosity effect on P700+ reduction; this result suggests a role for proton transfer in the PSI electron-transfer reactions. Second, we find that changes in hydration alter the rate of P700+ reduction and the interactions of P700 with the protein environment. This result suggests a role for bound water in electron transfer to P700+. Third, we present evidence that deuteration of the tyrosine aromatic side chain perturbs the vibrational spectrum, associated with P700+ reduction. We attribute this result to a linkage between CH-pi interactions and electron transfer to P700+.     

Total synthesis of (+)-amphidinolide A. Structure elucidation and completion of the synthesis

The structure elucidation of (+)-amphidinolide A, a cytotoxic macrolide, has been accomplished by employing a combination of NMR chemical shift analysis and total synthesis. The 20-membered ring of amphidinolide A was formed by a ruthenium-catalyzed alkene-alkyne coupling to forge the C15-C16 bond. Using the reported structure 1 as a starting point, a number of diastereomers of amphidinolide A were prepared. Deviations of the chemical shift of key protons in each isomer relative to the natural material were used as a guide to determine the locations of the errors in the relative stereochemistry. The spectroscopic data for the synthetic and natural material are in excellent agreement.     

Dynamic,computer-assisted interpretation of infrared spectra of condensed-phase mixtures

A knowledge-based system, MIXIR, designed to assist researchers in interpreting the infrared spectra of condensed-phase mixtures is described. This system overcomes many of the inherent limitations of static-rule-based systems. An iterative problem-solving approach is permitted, making use of information gained during the interpretation process. The dynamic rule selection and significance evaluation alos allow the user to provide information on the sample matrix, or to select a subset of the compounds for rule derivation. The MIXIR system was evaluated with a knowledge base compiled using IRBASE, a complementary program, and a test set of 20 mixtures.     

Synthesis of methyl-1,6-dioxaspiro[4,5]decanes using organoselenium mediated cyclization reactions

Three naturally occurring methyl-1,6-dioxaspiro[4.5]decanes have been prepared in good yield using organoselenium mediated reactions during the crucial cyclization process.     

Syntheses of seven-membered rings: ruthenium-catalyzed intramolecular [5+2] cycloadditions

The Ru-catalyzed intramolecular [5+2] cycloaddition of cyclopropylenynes is investigated with respect to the regio- and diastereoselectivity as well as the functional group compatibility of the reaction. Evidence for the mechanism as occurring through a ruthenacyclopentene intermediate is elucidated from 1) the study of the diastereoselectivity of the cycloaddition; 2) the effect of variation of substituents on the regioselectivity of cyclopropyl bond cleavage in 1,2-trans- and 1,2-cis-disubstituted cyclopropanes and 3) examples that clearly do not involve ruthenacyclohexene as intermediates as products still incorporate the cyclopropyl moiety. The scope and limitations of the Ru-catalyzed cycloaddition are discussed and compared with the Rh-catalyzed reaction. The potential power of this methodology towards natural product total synthesis is demonstrated by the formation of several polycyclic systems with the chosen reaction conditions and readily available cyclopropylenyne substrates.     

Polytriazoles as copper(I)-stabilizing ligands in catalysis

Polytriazolylamines were synthesized by the copper(I)-catalyzed ligation of azides and alkynes. The C3-symmetric derivative, TBTA, was shown to be a powerful stabilizing ligand for copper(I), protecting it from oxidation and disproportionation, while enhancing its catalytic activity.     

A chemoselective reduction of alkynes to (E)-alkenes

The trans reduction of all types of alkynes to give (E)-olefins is achieved through a two-stage trans hydrosilylation and protodesilylation. Reaction of an alkyne and a silane with the ruthenium catalyst [Cp*Ru(MeCN)3]PF6 results in clean hydrosilylation to give only the (Z)-trans addition product at ambient temperature with catalyst loadings of 1-5 mol %. The crude vinylsilane products are then protodesilylated by the action of cuprous iodide and TBAF at rt-35 degrees C. The reaction is compatible with many sensitive functional groups and provides a general trans-alkyne reduction not possible by other means.     

Fluorescence lifetimes of diphenylhexatriene-containing probes reflect local probe concentrations: Application to the measurement of membrane fusion

An important process in the life of a cell is fusion between cellular membranes. This is the process by which two cellular compartments surrounded by different membranes join to become a single compartment surrounded by a single membrane, without significant loss of compartment contents. To demonstrate fusion, the cell biophysicist must demonstrate all three critical aspects of the process: (1) mixing of membrane components, (2) mixing of compartment contents; and (3) retention of compartment contents. Most commonly, accomplishing this involves the use of fluorescence probes. The general theme to the methods described involves some form of concentration-dependent quenching. An unique method developed in our laboratory utilizes the concentration dependence of the fluorescence lifetime of a phosphatidylcholine containing carboxyethyl diphenylhexatriene at position 2 and palmitic acid at position 1 of glycerol (DPHpPC). The fluorescence lifetime of this molecule and that of its parent fluorophore diphenylhexatriene (DPH) shorten dramatically as their two-dimensional concentrations in a membrane increase. This lifetime quenching can be described by dimer formation that reduces the symmetry of the DPH excited state. This phenomenon allows one to use the fluorescence lifetime to gain insight into the local concentration of probe in microscopic regions of a membrane. One application of this is in distinguishing lipid transfer between the outer leaflets of two contacting membrane bilayers from fusion between these membranes that leads to mixing of lipids in both the inner and outer leaflets of the membrane bilayers. This allows a single measurement to demonstrate fusion between membrane pairs.Abbreviations PEG poly(ethylene glycol) - Na₂EDTA ethyiene-diamine-tetraacedic acid, disodium salt - LUV large, unilamellar vesicles made by rapid extrusion technique - DPH 1,6-diphenyl-trans-1,3,5-hexatriene - DPHpPC 1-palmitoyl-2-[[[2-[4- (phenyl-trans-1,3,5-hexatrienyl)phenyl]ethyl]oxy]carbonyl]-3-sn-phosphatidylcholine - DPPC 1,2-dipalmitoyl-3-sn-phosphatidylcholine - PA palmitic acid - NBD-PE N-(7-nitro-2,1,3-benzoxadiazol-4-yl)-PE - Rh-PE N-(lissamine Rhodamine B sulfoyl)-PE - R₁₈ octadecyl Rhodamine B chloride - ANTS 1-aminonaphthalene-3,6,8-trisulfonic acid - DPX N,N-p-xylylene-bis(pyradinium bromide)     

Operators with singular continuous spectrum,VII. Examples with borderline time decay

We construct one-dimensional potentialsV(x) so that if onL ²(), thenH has purely singular spectrum; but for a dense setD, D implies that |,e ^-itH |C |t|^-1/2 ln(|t|) for t>2. This implies the spectral measures have Hausdorff dimension one and also, following an idea of Malozemov-Molchanov, provides counterexamples to the direct extension of the theorem of Simon-Spencer on one-dimensional infinity high barriers.This material is based upon work supported by the National Science Foundation under Grant No. DMS-9401491. The Government has certain rights in this material.