Pyrene. Probe of lateral diffusion in the hydrophobic region of membranes

Vanderkooi, Jane M.; Callis, James B.

doi:10.1021/bi00716a028

Cited by 299 publications

(143 citation statements)

References 32 publications

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“…The fluorescence emission intensity of the excimer peak of pyrene (470 nm) can be conveniently used to monitor membrane fluidity (30,41), since the formation of the excimer is related to the lateral mobility of pyrene molecules in the lipid phase (6,40).…”

Section: Discussionmentioning

confidence: 99%

The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain: purification, structure, and interaction with synthetic membranes

1991

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The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain was purified to homogeneity from methanol extracts of dried cells by reverse-phase liquid chromatography and was designated P-3. On the basis of the UV-visible, infrared, mass, and 'H nuclear magnetic resonance spectra of P-3, it was identified as bisdehydro-,-carotene-2-carboxylic acid. The pigment interacted with synthetic membranes of phosphatidylcholine and dimyristoyl phosphatidylcholine and stabilized the membranes. These results also indicate that P-3 is different from canthaxanthin, the major carotenoid pigment from a mesophilic M. roseus strain.Carotenoid pigments are present in a wide variety of bacteria, algae, fungi, and plants (1,23 (16,17). Cells were pelleted by centrifugation at 1,000 x g for 10 min, washed free of medium with distilled water, and freeze-dried to a powder. Methanol (100 ml) was added to about 1 g of freeze-dried cells, and the mixture was vortexed until the methanol layer turned red. The entire suspension of cells was centrifuged (5,000 x g for 5 min), and the methanol layer containing the crude pigment was recovered. The pale pink 7911 JOURNAL

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Section: Discussionmentioning

confidence: 99%

The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain: purification, structure, and interaction with synthetic membranes

1991

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“…In spite of the fact that the fluorescence probes used are distributed differently in a membrane, DMC and MBA being closer to the membrane-water interface [18,19] and pyrene [21] and perylene [22] -in the depth of lipid bilayer, they have provided similar values of the increase in membrane viscosity after lipid peroxidation.…”

Section: Discussionmentioning

confidence: 99%

The increase of phospholipid bilayer rigidity after lipid peroxidation

Dobretsov¹,

Borschevskaya²,

Петров³

et al. 1977

FEBS Letters

276

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“…As can be seen, a large increase in the emission intensity of the excimer relative to that of the monomer occurs as a result of treatment with colicin Ia. The phenomenon of excimer formation is well understood (14,15). The transient excimer species forms upon collision of a molecule in the first singlet excited state and a molecule in the ground state.…”

Section: Methodsmentioning

confidence: 99%

Affinity of intact Escherichia coli for hydrophobic membrane probes is a function of the physiological state of the cells.

Nieva-Gomez¹,

Gennis²

1977

Proc. Natl. Acad. Sci. U.S.A.

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The fluorescence parameters of several common membrane probes in the presence of whole E. coli have been examined. The probes included electrically neutral lipophilic molecules N-phenyl-1-naphthylamine, pyrene, and 1,6-diphenyl-1,3,5-hexatriene as well as the negatively charged molecule 8-anilino-1-naphthalene sulfonate. It is demonstrated in each case that certain fluorescence parameters are a function of the state of energization of the cells.All the probes appear to monitor structural changes in the E. coli envelope which accompany the energization and de-energization of the cells. The use of fluorescent probes to study membrane structure and processes has become widespread. Electrically charged probes such as 8-anilino-1-naphthalene sulfonate (ANS) and cyanin dyes have been used to monitor membrane potential in liposomes and membrane preparations from mitochondria and bacteria (1-3). Uncharged lipophilic probes such as N-phenyl-1-naphthylamine (NPN), pyrene, and 1,6-diphenyl-1,3,5-hexatriene (DPH) have been used primarily to monitor the fluidity or microviscosity of the lipid bilayer (4-6). It has recently been shown that the fluorescence parameters of NPN in the presence of whole Escherichia coli cells are a function of the physiological state of the cells (7). Cells that are de-energized bind significantly more NPN than cells that are energized, leading to about a 4-fold increase in the fluorescence intensity when the cells are de-energized by such treatments as substrate or oxygen depletion, addition of electron transport inhibitors, addition of uncouplers, or addition of colicin Ia. The fluorescence changes can be rapidly reversed if the cells are restored to the energized state by addition of oxygen to anoxic cells or the addition of substrate to starved cells. It appears that this lipophilic probe monitors structural changes in the Escherichia coli envelope which occur reversibly as the cells are energized and de-energized. These results probably explain at least in part the changes in NPN fluorescence observed with colicin-El-treated cells (8). The effects of these colicins on the cells resemble in several respects the effects of uncouplers (9, 10).In an attempt to determine the nature of these structural changes in the cell envelope, the present work reports studies with probes that have fluorescent, structural, and electrical properties different from those of NPN. In addition, a photoreactive probe, 1-azidopyrene, has been used to confirm the conclusion that there is a reversible change in the affinity of the cells for hydrophobic molecules as the cells are energized and de-energized. Upon irradiation, a highly reactive nitrene is generated (11), resulting in covalent, fluorescent adducts presumably with components of the bacterial envelope. MATERIALS AND METHODSCell Growth. E. coli K-12 strain JK1 was grown at 370 in M9 medium, which consists of glucose at 1.3 g/liter and M9 salts (M9S). M9S is composed of the following (in g/liter): NH4Cl, 1.0; MgSO4, 0.13; KH2PO4, 3.0; Na2HPO4, 6.0. Cell...

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Pyrene. Probe of lateral diffusion in the hydrophobic region of membranes

Cited by 299 publications

References 32 publications

The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain: purification, structure, and interaction with synthetic membranes

The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain: purification, structure, and interaction with synthetic membranes

The increase of phospholipid bilayer rigidity after lipid peroxidation

Affinity of intact Escherichia coli for hydrophobic membrane probes is a function of the physiological state of the cells.

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