Nitrite-reducing activity of modified cytochrome c-553 from the red alga Porphyra yezoensis Ueda

Crystalline cytochrome c-553 was obtained from Porphyra yezoensisUeda. The cytochrome in areduced form was modified to show anitrite-reducing activity after appropriate treatment with heat,hydrogen peroxide, or photooxidation using methylene blue asthe electron acceptor, but the reducing activity was far lowerthan that of the nitrite reductase isolated from this alga.The modified cytochrome c-553 was autooxidizable and showedan absorption spectrum resembling that of cytochrome c-553 inthe oxidized form except for slight shifts of the absorptionmaximumin the -band region toward shorter wavelengths. ¹ Present address: Department of Biological Sciences, Universityof Tsukuba, Sakura-Mura, Ibaraki, 300-31 Japan. ² Present address: Department of Fisheries, College of Agricultureand Veterinary Medicine, Nihon University, Shimouma, Setagaya-ku,Tokyo, 154 Japan. (Received June 10, 1975; )     

Nitrobacter agilis Cytochrome c-550: Isolation, Physicochemical and Enzymatic Properties, and Primary Structure

Nitrobacter agilis cytochrome c-550 was purified to an electrophoreticallyhomogeneous state, and some of its properties were determined.The cytochrome showed an absorption peak at 410 nm in the oxidizedform, and peaks at 416, 521 and 550 nm in the reduced form.Its isoelectric point was 8.1 at 5?C. Analysis of the aminoacid composition showed that the cytochrome molecule was composedof 108 amino acid residues, 16 of which were lysine residues. The cytochrome reacted rapidly with N. agilis cytochrome c oxidaseand yeast cytochrome c peroxidase and more slowly with Pseudomonasaeruginosa nitrite reductase and bovine cytochrome c oxidase.The reactivities with these redox enzymes suggested that thecytochrome might be an evolutionary stage between bacterialand eukaryotic cytochromes c. The primary structure of the cytochrome from the N-terminusto the 85th residue was determined. The N-terminal sequencewas homologous to the corresponding portion of the primary structureof horse cytochrome c. ¹ Present adress: Department of Chemistry, Faculty of Science,Tokyo Institute of Technology, O-okayama, Meguro-ku, Tokyo,152, Japan. (Received December 3, 1981; Accepted January 28, 1982)     

A probable lack of function of c-type cytochrome in the photosynthetic system of the blue-green alga Anabaena variabilis

Quantitative study of the cytochrome c acting in the photosyntheticsystem of the blue-green alga Anabaena variabilis (M-2) wasdone with membrane fragments and intact cells. Membrane fragments highly active in the NADP⁺-Hill reaction(above 200 µmoles/mg chl.a;-hr) retained photoresponsivecytochrome c equal only one-tenth that of P700, while the plastocyanincontent was almost equal to that of P700. The cytochrome contentin intact cells was a little larger than that in membrane fragmentson the chlorophyll a basis. However, the values relative toP700 (1/9) and plastocyanin (1/10) were identical with thosein membrane fragments. The content was also far smaller thanthat of reaction center II's (1/6). If the cytochrome mediatesall electrons from reaction center II, the cytochrome oxidation-reductionshould have a rate constant of 2.4?102 sec^–1 which isone order above of the rate constant of the cytochrome reduction(2.3 to 3.5?10¹sec^–1). These quantitative relationshipsindicate that in Anabaena variabilis (M-2), c-type cytochrome,either cytochrome f or algal cytochrome c, cannot function inthe main electron flow between two reaction centers. (Received September 8, 1978; )     

Mechanisms of the acido- and thermophily of Cyanidium caldarium Geitler V. Acid and heat stabilities of soluble proteins

Acid and heat stabilities of water-soluble proteins extractedfrom an acidophilic and thermophilic unicellular alga, Cyanidiumcaldarium, by French pressure cell treatment were investigated.Soluble proteins from Cyanidium were not particularly acid orheat stable compared with those from mesophilic algae such asAnabaena variabilis and Anacystis nidulans. The results suggestthat the acidophilic and thermophilic mechanisms of Cyanidiumcan not be ascribed to the acid and heat stabilities of theproteins in the cells. ¹ Based on a dissertation submitted to the Tokyo MetropolitanUniversity in partial fulfillment of the requirements for thedegree of Doctor of Science. (Received October 24, 1977; )     

Studies on cytochromes in photosynthetic electron transport system I. photoreduction and photo-oxidation of cytochrome b559 by photosystem II in spinach chloroplasts

Light-induced redox-reactions of cytochrome b₅₅₉ in spinachchloroplasts were investigated. Illumination of chloroplastsinduced photoreduction of cytochrorne b₅₅₉ Red light (650 nm)was more effective than far-red light (725 nm), indicating thatthe photoreduction is a photosystem II-mediated reaction. Onaddition of DCMU, the photoreduction was eliminated and a photooxidationof cytochrome b₅₅₉ was observed. The rate of this photooxidationwas faster with photosystem II light than with photo-systemI light. On addition of Mn⁺⁺ the photooxidation was partly suppressed;far-red light became as effective as red light in inducing photooxidationof cytochrome b₅₉₉, in the presence of DCMU and Mn⁺⁺. Ascorbate completely suppressed photooxidation of cytochromeb⁵⁵⁹ In the presence of ascorbate, however, photooxidation wasobserved in the presence of inhibitors or after inhibitory treatmentsof chloroplasts which affected the oxidizing side of systemII. These inhibitors and inhibitory treatments, but not DCMU,decreased the redoxpotential of cytochrome b₅₅₉. Reactivationof Hill reaction in Tris-washed chloroplasts by indophenol-ascorbatetreatment was not accompanied by an abolishment of photooxidationof cytochrome b₅₅₉. A possible mechanism is proposed to account for these reactionsof cytochrome b₅₅₉ in the photosynthetic electron transportin chloroplasts. (Received April 4, 1972; )     

Salicylic Acid Levels in Thermogenic and Non-Thermogenic Plants

The natural trigger for heat production in the thermogenic inflorescencesof Sauromatum guttatum Schott (voodoo lily) was recently identifiedas salicylic acid (SA), which induced heat production at levelsas low as 13 ng g f. wt^–1. Since then the levels of SAwere determined in other thermogenic and non-thermogenic plantspecies. In thermogenic inflorescences of five aroid species,and in male cones of at least four thermogenic cycads SA levelsduring heat production exceeded 1 µg g f. wt^–1.SA was not detected in the thermogenic flowers of a water lily,Victoria regia Lindl. (Nymphaeaceae), and Bactris major Jacq.(Palmae). Levels of salicylic acid varied substantially in thefloral parts of seven non-thermogenic species and in the leavesof 27 non-thermogenic species. Amorphophallus campanulatus Blume ex Decne, Arum italicum Mill., Arum dioscoridis Sibth. & Son., Philodendron selloum Koch, Monstera deliciosa Liebm., Encephalartosferox Bertol. f., Encephalartos hildebrandtii A. Br. & Bouché, Encephalartos gratus Prain, Dioon edule Lindl. cv. edule, Dioon edule Lindl. cv angustifolium, Sauromatum guttatum Schott, voodoo lily, Victoria regia Lindl., Bactris major Jack, salicylic acid, thermogenicity, heat production     

CHANGE OF B-TYPE HAEM CONTENT IN RELATION TO PEROXIDASE BIOSYNTHESIS IN INJURED SWEET POTATO ROOTS

The methods of quantitative analysis of b-type haem in plantswere investigated. With an improved method developed was determinedthe haem content in the supernatant, mitochondrial, and microsomalfractions of sweet potato tissue. The activities of peroxidase,catalase, and cytochrome oxidase, as well as the contents ofb-type haem and acid-insoluble nitrogen in the cellular fractionswere determined at different incubation times after cuttingof sweet potato tissue. Peroxidase and catalase increased withtime in each celluler fraction, following a short lag phase.In the mitochondrial fraction, b-type haem, cytochrome oxidase,and acid insoluble nitrogen increased linearly with time. Inthe microsomal and supernatant fraction, b-type haem increasedwith time following a short lag phase. The increase in haemcontent of the supernatant fraction appeared to be associatedwith peroxidase formation. Time course analysis showed that ⁵⁹Fe-incorporation into b-typehaem of the supernatant fraction increased with time and thatincorporation was markedly inhibited by blasticidin S. The incorporationof ⁵⁹Fe into mitochondrial haem did not increase with time andwas not inhibited by blasticidin S. Blasticidin S inhibited⁵⁹Fe-incorporation into microsomal haem. Time course analysisof b-type haem content, ⁵⁹Fe-incorporation into b-type haem,and peroxidase activity suggest that in the injured tissue haemis synthesized from low molecular weight compounds and is incorporatedinto peroxidase as the haem moiety. ¹ This paper constitutes Part 57 of the Phytopathological Chemistryof Sweet Potato with Black Rot. ² Present address: Institute for Plant Virus Research, Chiba.     

COMPONENTS OF THE ELECTRON-TRANSFERRING SYSTEM IN ACETOBACTER SUBOXYDANS AND RECONSTRUCTION OF THE LACTATE OXIDATION SYSTEM

1. Cytochromes a₁⁵⁹⁰, b⁵⁶⁰, c₁⁵⁵⁴ and c₁⁵⁵² were isolated andpurifiedfrom a strain of Acetobacter suboxydans. The proceduresusedwere described in detail.
2. The main cytochrome band at550-560 mµ in intact cellssplitted at liquid air temperatureinto two bands, 551 mµ(strong) and 559 mµ (weak).
3. Optical and physiological properties of the four cytochromeswere investigated. Lactic dehydrogenase activity was found tobe associated with cytochrome c₁⁵⁵⁴. The two c₁-type cytochromes,especially cytochrome c₁⁵⁵⁴, persisted in their reduced formafter the purification through many steps.
4. By some combinationsof isolated components reconstruction ofthe oxygen uptake systemcould be realized.
5. The oxygen-consuming activity of purifiedoxidase preparationswas accelerated by a-tocopherol but notby Emasoll 4130 andTween 80.
6. Some discussions were made onthe nature of terminal oxidase,the role of cytochrome c₁⁵⁵²in the electron-transport system,and persistence of reducedstate of c₁-type cytochromes.
7. A possible scheme of the electron-transferringsystem of Acetobactersuboxydans was presented.

(Received May 16, 1960; )     

Studies on the metabolism of a sulfur-oxidizing bacterium VII. Oxidation of sulfite by a cell-free extract of Thiobacillus thiooxidans

Properties of the cell-free extract, prepared from a strainof Thiobacillus thiooxidans by sonic disruption followed byfractionation with centrifugatiori, were investigated with referenceto its sulfite-oxidizing activity. Without the addition of cofactors the particulate fraction(F-P)catalyzed oxidation of sulfite with oxygen or bacterial cytochromec-552 obtained from Pseudomonas stutzeri as electron acceptor.TMPD reduced by ascorbic acid was also oxidized by F-P. Thesoluble fraction(F-S) showed no activity in oxidizing sulfiteand TMPD, but stimulated TMPD oxidation by F-P. Oxygen uptake with either sulfite or TMPD as substrate was inhibitedby KCN, NaN₃, CO and c-phenanthroline. CO-Inhibition was reversedby light. Reduction of cytochrome c-552 by sulfite was insensitiveto these agents. Antimycin A markedly inhibited sulfite oxidation with eitheroxygen or cytochrome c-552 as electron acceptor, but was withouteffect on TMPD oxidation. DDC and SAO, both strong inhibitors of sulfur oxidation, didnot affect sulfite and TMPD oxidations. Cytochromes of the a, b and c types were contained in F-P. Thesecytochromes were rapidly reduced when F-P was incubated withsulfite. Cytochrome(s) of the c type was present in F-S, too. ¹VI.=References (3) ²Partly supported by a grant from the Ministry of Education ³Present address: Sanyo Women's College, Hatsukaichi, Hiroshima738, Japan ⁴Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima 734, Japan (Received May 15, 1970; )     

Cytochrome components of green alga, Bryopsis maxima: Purification and properties of cytochrome f from membrane fragments

From the membrane fragments of the green alga Bryopsis maxima,a cytochrome which resembles cytochrome f of higher plants wassolubilized with methyl ethyl ketone. The cytochrome was partlypurified by ammonium sulfate fractionation, followed by gelfiltration. Its properties were similar to those of the algalcytochrome f reported by Wood (26). The approximate molar ratioof cytochromes f, c-553 and chlorophyll in B. maxima was 1 :1 : 600–700. ¹ In this communication, according to the recommendation byWood (26), cytochrome f is the membrane-bound c-component andcytochrome c-55 the soluble one. In some references cited, thesechloroplast cytochromes are called algal cytochrome f. (Received February 16, 1978; )     

Contents of Cytochromes, Quinones and Reaction Centers of Photosystems I and II in a Cyanobacterium Synechococcus sp.

The contents of photosystem I and photosystem II reaction centers,cytochrome c-553, cytochrome c-550, cytochrome f, cytochromeb-559, cytochrome b-563, plastoquinone and vitamin K₁ in thecyanobacterium Synechococcus sp. were determined. About threephotosystem I reaction centers were present for each photosystemII reaction center. The amounts of cytochromes functioning betweenthe two photosystems were approximately half those of the photosystemI reaction center. Plastocyanin was not detected, while plastoquinoneand vitamin K₁ were present in excess of other electron carriersand reaction centers. The results indicate the importance ofplastoquinone and cytochrome c-553 for cooperation of the tworeaction centers through electron transport. ¹Present address: Toray Basic Research Laboratory, 1111 Tebiro,Kamakura, Kanagawa 248, Japan. (Received June 17, 1982; Accepted January 17, 1983)     

Intra- and extracellular measurement of reactive oxygen species produced during heat stress in diaphragm muscle

Skeletalmuscles are exposed to increased temperatures during intense exercise,particularly in high environmental temperatures. We hypothesized thatheat may directly stimulate the reactive oxygen species (ROS) formationin diaphragm (one kind of skeletal muscle) and thus potentially play arole in contractile and metabolic activity. Laser scan confocalmicroscopy was used to study the conversion of hydroethidine (a probefor intracellular ROS) to ethidium (ET) in mouse diaphragm. During a30-min period, heat (42°C) increased ET fluorescence by 24 ± 4%, whereas in control (37°C), fluorescence decreased by 8 ± 1% compared with baseline (P < 0.001). The superoxidescavenger Tiron (10 mM) abolished the rise in intracellularfluorescence, whereas extracellular superoxide dismutase (SOD; 5,000 U/ml) had no significant effect. Reduction of oxidized cytochromec was used to detect extracellular ROS in rat diaphragm.After 45 min, 53 ± 7 nmol cytochrome c · g drywt¹ · ml¹ were reduced in heatcompared with 22 ± 13 nmol · g¹ · ml¹ in controls(P < 0.001). SOD decreased cytochrome creduction in heat to control levels. The results suggest that heatstress stimulates intracellular and extracellular superoxideproduction, which may contribute to the physiological responses tosevere exercise or the pathology of heat shock.

     

Purification and properties of cytochrome f from Brassica komatsuna leaves

Cytochrome f was extracted from leaves of Brassica komatsuna(Brassica Rapa L. var. pervidis Bailey) in an aqueous solutionusing methyl ethyl ketone and was purified by the followingsteps: (i) acetone precipitation, (ii) ammonium sulfate fractionation(0.33–0.7 saturation), (iii) DEAE-cellulose column chromatography,and (iv) Sephadex G-100 column chromatography. Characteristic spectroscopic properties and the midpoint potentialof the cytochrome were essentially identical with those of thecytochrome f from parsley reported by Bendall et al. Molecular weight of the cytochrome determined by gel filtrationwas close to 32,000 and it contained one haem per molecule ofprotein. The ferro-cytochrome was oxidized by potato polyphenol oxidasein the presence of chlorogenic acid. Under light-aerobic conditions, the ferro-cytochrome was rapidlyoxidized by the chlorophyll-protein CP743 from Chenopodium albumin the presence of menadione. Under light-anaerobic conditions,the oxidized cytochrome was reduced at a considerable rate. ¹ Cytochrome c₆ according to the enzyme nomenclature recommendedby I.U.P.A.C.-I.U.B. (5). (Received November 7, 1974; )     

COMPARATIVE STUDIES OF PHOTOCHEMICAL OXIDATION-REDUCTION REACTIONS IN LAMELLAR FRAGMENTS OF VARIOUS ALGAE AND SPINACH

The activity of various electron carriers, including DPIP, spinachplastocyanin, mammalian cytochrome c, and Anabaena cytochrome553, as donor in the reaction induced by the photochemical systemI was examined with lamellar fragments of various algae andspinach. Reduced DPIP was an effective electron donor irrespective ofthe organisms, when it was supplied at a high concentration(10^–3 M). Spinach plastocyanin was effective in the reactionswith the lamellae of green algae, Euglena, diatom Phaeodactyrumand red algae Porphyra yezoensis and Porphyra sp. Yamamoto II,whereas it was inactive in the lamellae of blue-green algae.Horse-heart cytochrome c and Anabaena cytochrome 553 were activein the reaction with the lamellae of bluegreen algae. The formercytochrome was also active in the reactions in Porphyridiumand Cyanidium. The cytochromes were less active in the reactionsin which spinach plastocyanin acted as effective electron donor. The data were interpreted as that the photochemical system Iin bluegreen algae differs from that of other photosyntheticorganisms with respect to the properties of the site of theelectron-input. ¹ Present address: Nomura Research Institute for Technologyand Economics, Kamakura, Kanagawa. ² Present address: Ocean Research Institute, University of Tokyo,Nakano, Tokyo.     

Tetcyclacis and Abscisic Acid-Sensitive Reduction of Extracellular Ferricyanide by Mesophyll Cells of Valerianella locusta and Lemna gibba

Electron transport across the plasma membrane of Valerianellalocusta mesophyll cells and intact fronds of Lemna gibba, inducedby 10^–3 M ferricyanide, was inhibited by tetcyclacis,an inhibitor regarded to be specific for cytochrome P-450 mono-oxygenases.The effect was dependent on the concentration of tetcyclacisand the duration of preincubation. The apparent rate of trans-membraneelectron transport increased in the presence of catalase, indicatingtetcyclacis-induced H₂O₂-production or additional tetcyclacis-independentH₂O₂ release. The findings suggest an interaction of cytochromeP-450 with the plasma membrane-located electron transport chain.This redox-chain could be involved in the degradation of abscisicacid, being located at the plasma membrane. This assumptionis supported by the finding that ABA inhibits extracellularferricyanide reduction. Key words: Abscisic acid, cytochrome P-450 mono-oxygenase, plasma membrane, tetcyclacis     

Purification of Sulphite-Cytochrome c Reductase of Thiobacillus novellus and Reconstitution of Its Sulphite Oxidase System with the Purified Constituents

Sulphite-cytochrome c reductase (sulphite: ferricytochrome coxidoreductase, EC 1.8.2.1 [EC] ) derived from Thiobacillus novelluswas purified by chromatography on a DEAE-cellulose column andby gel filtration with a Sephadex G-100 column. Although thereductase thus purified moved as a single band both in gel filtrationand in isoelectric focusing it was always split into two bandsby polyacrylamide gel electrophoresis; the one had the enzymaticactivity and showed absorption spectrum of cytochrome, whilethe other had no activity and was colourless, in contrast withthe results reported by Charles and Suzuki [(1966) Biochim.Biophys. Acta 128: 522]. The enzymatic properties of the purifiedreductase were almost the same as those of the enzyme obtainedby Charles and Suzuki. Cytochrome c-551 free of the reductase activity was obtained.Its molecular weight was determined to be 23,000 by polyacrylamidegel electrophoresis in the presence of sodium dodecyl sulphate.The cytochrome seemed to exist in the organism as a complexwith the reductase or a subunit of the enzyme. In the stateof the complex with the enzyme, the cytochrome was reduced veryquickly on addition of sulphite, while the cytochrome free ofthe reductase activity was hardly reduced by the enzyme withsulphite. A sulphite oxidase system was reconstituted with the reductase,cytochrome c-550 and cytochrome oxidase highly purified fromthe bacterium. ¹ Present address: Water Research Institute, Nagoya University,Nagoya 464, Japan ² Present address: Institute for Biological Science, SumitomoChemical Co., Ltd., Takarazuka, Hyogo 665, Japan (Received January 23, 1981; Accepted March 9, 1981)     

Different Intracellular Localization of Two Cytochrome P-450 Systems, Ipomeamarone 15-Hydroxylase and Cinnamic Acid 4-Hydroxylase, in Sweet Potato Root Tissue Infected with Ceratocystis fimbriata

Ipomeamarone 15-hydroxylase activity was mainly recovered inthe pellet fraction between centrifugations at 10,000 and 100,000?gfrom a crude extract of Ceratocystis fimbriata-infected sweetpotato root tissue, whereas cinnamic acid 4-hydroxylase activitywas found between centrifugations at 300 and 10,000?g. Whenparticles in the crude extract were fractionated by sucrosedensity gradient centrifugation, the rough-surfaced microsomeswere distributed over a wide density range from 1.09 to 1.14g cm^–3, judging from the distributions of protein, RNAand NADPH-cytochrome c reductase activity. Phosphorylcholine-glyceridetransferase activity was only in the lighter half of the microsomalfraction (density: 1.09–1.11 g cm^–3). Ipomeamarone15-hydroxylase activity was found in heavier half of the microsomalfraction (density: 1.10–1.14 g cm^–3). We proposethat this tissue has two rough-surfaced endoplasmic reticulumspecies, only one of which carries phosphorylcholine-glyceridetransferase, and that the cytochrome P-450 system is localizedon the species lacking the enzyme. Cinnamic acid 4-hydroxylaseactivity was mainly found in a fraction that had densities of1.17–1.19 g cm^–3 and contained vesicular particlesof various sizes. ¹ Present address: Laboratory of Food Hygienics, Faculty ofAgriculture, Kagawa University, Miki-cho, Kida-gun, Kagawa 761-07,Japan. (Received September 6, 1984; Accepted December 27, 1984)     

Oxidation Rate of Monomeric Radish Cytochrome f with Plastocyanin

The reaction rate of reduced monomeric cytochrome f with oxidizedplastocyanin, both purified from Japanese radish, was determinedby a stopped-flow method. The oxidation rate constant was 6.0x 10⁷ M¹sec¹ at pH 7.0 and 25°C, which is slightly higherthan the value reported by Wood [(1974) Biochim. Biophys. Acta357 : 370] for oligomeric parsley cytochrome f Thermodynamicparameters also were determined to be 56 KJ M^–1 for activationenthalpy and 90 J M^–1 K^–1 for activation entropy.Neither a pH from 6 to 9 nor the addition of NaCl, polylysine,histone or polyaspartate affected the rate constant. ¹Present address: The National Institute for Environmental Studies,Yatabe, Ibaraki 305, Japan. (Received October 9, 1980; Accepted November 17, 1980)     

The Oxidative Phosphorylation and ATPase Activity of Arum spadix Mitochondria in Relation to Heat Generation

The respiration of Arum spadix mitochondria is coupled to asub-maximal stoichiometry of ATP synthesis. The P/O ratios associatedwith the oxidation of succinate or malate are decreased by antimycinand increased by m-chlorobenzhydroxamic acid, an inhibitor ofthe alternative oxidase. The mitochondrial ATPase activity of20–40 nmol (mg protein)^–1 min^–1 is independentof the maturity of the spadix and is unlikely to provide themechanism for heat production during the odoriferous stage,which probably results from an increase in the rate of electrontransport via the non-phosphorylating, cyanide-insensitive oxidase.     

Multiple eicosanoid-activated nonselective cation channels regulate B-lymphocyte adhesion to integrin ligands

Arachidonic acid (AA) is a substrate for a variety of proinflammatory mediators, which are generated by cyclooxygenases (COXs), lipoxygenases (LOXs), and cytochrome P-450 (CYP450) enzymes. COX (e.g., PGs and prostacyclins) and LOX (e.g., leukotrienes) products have well-established proinflammatory roles; however, little is known about the functions of CYP450 products in leukocytes. We previously found that mechanical strain generated by subjecting lymphocytes to hypotonic challenge triggered AA production and that two CYP450 products of AA, 5,6-epoxyeicosatrienoic acid (5,6-EET) and 20-hydroxyeicosatetraenoic acid (20-HETE), as well as a product of LOX, 5-(S)-hydroperoxyeicosatetrenoic acid (5-HPETE), induced Ca²⁺ entry into primary B cells. The main goal of the present studies, therefore, was to define the biophysically properties of eicosanoid-activated channels responsible for Ca²⁺ entry and the physiological consequences of activating these channels, including their role in mechanical signaling. We found that 5,6-EET, 20-HETE, and 5-HPETE each activated distinct Ca²⁺-permeant nonselective cation channels (NSCCs) in primary B cells. These NSCCs each regulate plasma membrane potential and B-cell adhesion to integrin ligands ICAM-1 and VCAM-1. Thus our data demonstrate that proinflammatory mediators produced in response to osmotic and/or physical stress play a direct role in regulating the B-cell membrane potential and their adhesion to specific ECM proteins. These results not only have important implications for understanding normal mechanisms of B-cell activation, differentiation, and trafficking but also point to novel targets for modulating the pathogenesis of B-cell-mediated inflammatory diseases. calcium; arachidonic acid; membrane potential; hypotonicity; cytochrome P-450