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T Ahn

Publications and source records attributed to T Ahn.

At least 19 recordsLinked to original sources
PubMed2001

Effects of lipids on the interaction of SecA with model membranes.

The effects of nonlamellar-prone lipids, diacylglycerol and phosphatidylethanolamine (PE), on the kinetic association of SecA with model membranes were examined by measuring changes in the intrinsic emission fluorescence with a stopped-flow apparatus. Upon interaction with standard liposomes composed of 50 mol% dioleolyphosphatidylcholine (DOPC) and 50 mol% of dioleoylphosphatidylglycerol (DOPG), the intrinsic fluorescence intensity of SecA was decreased after a lapse of time with a rate constant of 0.0049 s(-1). When the DOPC of the standard vesicles was gradually replaced with either dioeloyl PE (DOPE) or Escherichia coli (E. coli) PE, the rate constant increased appreciably as a function of PE concentration, in the order DOPE > E. coli PE. In addition, when the PE of E. coli PE/DOPG (50/50) vesicles was replaced with more than 5 mol% dioleoylglycerol (DOG), the rate constant further increased by 40%. The incorporation of nonlamellar-prone lipids in the vesicles also enhanced the binding of SecA to model membranes in the order DOPE > or = E. coli PE/DOG > E. coli PE > DOPC. These results provide the first kinetic evidence for the importance of nonlamellar-prone phospholipids for the association rate of SecA with membranes.

Adenosine Triphosphatases↗
PubMed2001

Effect of divalent cations on the ATPase activity of Escherichia coli SecA.

It was found that Ca(2+) stimulates the intrinsic SecA ATPase activity in the absence as well as in the presence of liposome. On the other hand, Mg(2+), the general cofactor for ATPase, did not affect the intrinsic SecA ATPase but reduced the portion of ATPase activity enhanced by Ca(2+). The enhancement of SecA ATPase activity correlated well with the increase in 8-anilino-1-naphthalene-sulfonic acid binding of SecA, suggesting that increased exposure of hydrophobic residues stimulates the enzyme activity.

Adenosine Triphosphatases↗
PubMed2001

Microscopic polyangiitis complicated with massive intestinal bleeding.

Microscopic polyangiitis (MPA) is associated with renal dysfunction, in most cases, and occasionally with pulmonary hemorrhage. However, massive intestinal bleeding is a rare manifestation. We report a case of MPA in a man who developed arterial bleeding in the small intestine. A 74-year-old man was admitted after enduring a fever for 4 weeks. Laboratory examination revealed leucocytosis with neutrophil predominance, and renal dysfunction was noted. He did not respond to treatment with antibiotics, and the worsened renal function soon required hemodialysis. On the seventh hospital day, he suddenly developed massive melena. A colonoscopic examination suggested bleeding in the small intestine, and abdominal angiography demonstrated that the bleeding originated in a branch of the ileal artery. Hemostasis was achieved by transarterial embolization (TAE), but the patient suffered a massive pulmonary hemorrhage 2 days after the TAE. Although he responded well to a combination treatment with corticosteroid and cyclophosphamide, recurrence of pulmonary hemorrhage led to death, on the 87th hospital day. MPA in this patient was associated with three serious complications; deteriorating renal function, massive melena, and pulmonary hemorrhage. Of the various manifestations associated with MPA, arterial bleeding in the gastrointestinal tract, although rare, should be considered as one of the serious complications in MPA.

Aged↗
PubMed2001

ORE9, an F-box protein that regulates leaf senescence in Arabidopsis.

Senescence is a sequence of biochemical and physiological events that constitute the final stage of development. The identification of genes that alter senescence has practical value and is helpful in revealing pathways that influence senescence. However, the genetic mechanisms of senescence are largely unknown. The leaf of the oresara9 (ore9) mutant of Arabidopsis exhibits increased longevity during age-dependent natural senescence by delaying the onset of various senescence symptoms. It also displays delayed senescence symptoms during hormone-modulated senescence. Map-based cloning of ORE9 identified a 693-amino acid polypeptide containing an F-box motif and 18 leucine-rich repeats. The F-box motif of ORE9 interacts with ASK1 (Arabidopsis Skp1-like 1), a component of the plant SCF complex. These results suggest that ORE9 functions to limit leaf longevity by removing, through ubiquitin-dependent proteolysis, target proteins that are required to delay the leaf senescence program in Arabidopsis.

Abscisic Acid↗
PubMed2001

Influence of stent expansion states on platelet deposition in an extracorporeal porcine arteriovenous shunt model using a multichannel perfusion chamber.

Limited data are available about incomplete stent expansion (SE) on platelet deposition (PD). We examined PD following different SE using an extracorporeal porcine arteriovenous shunt model to which a perfusion chamber with four parallel silastic tubes were connected. Blood flow was set at a 20 and 100 mL/min in 1.8 and 3.1 mm diameter tubes, respectively. P154 stents were deployed completely (Group A, n=15) or incompletely (Group B, n=15) in 1.8 mm (n=13) and 3.1 mm (n=17) tubes. 51Cr-labelled platelet autologous blood was injected 1 hr before the perfusion. After 15 min-perfusion, the testing tubes were assessed for radioactivity counts. In-stent cross sectional area was measured by intravascular ultrasound. There was a significant difference in PD between group A and B regardless of channel size (118+/-18.4 vs 261.4+/-52.1 pits x 10(6)/cm2, p<0.05). With adjusted shear rate and similar stenosis, PD was similar in both tubes. In smaller 1.8 mm tubes, a stenosis as subtle as 10% was associated with a significant PD difference (226.1+/-20 vs 112.9+/-20.5 plts x 10(6)/cm2, p<0.005). This model enabled a repetitive, simultaneous comparison of PD following different SE states. It seems that the quality of SE remains crucial in smaller channels.

Animals↗
PubMed2000

Importance of phosphatidylethanolamine for the interaction of apocytochrome c with model membranes containing phosphatidylserine.

The effect of phosphatidylethanolamine (PE) on the binding of apocytochrome c to model membranes was examined. When 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) of the standard vesicles composed of 80% of this lipid and 20% of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS) was gradually replaced with upward of 50% of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), the binding increased appreciably. Ca(2+), causing the phase separation of PS, also brought about increased binding of apocytochrome c in the PC/PS system, underlining the importance of PS properties in membranes for the protein binding. The resonance energy transfer between Trp-59 in apocytochrome c and pyrene-PS incorporated into bilayers showed that the replacement of PC with PE increased the extent of apocytochrome c penetration into membranes by a PE concentration-dependent manner. However, in the absence of PS, PE had no apparent effect on these functions of apocytochrome c, suggesting that PE-induced change(s) of acidic membrane properties is important to the association of apocytochrome c with vesicles. From the observations that the excimer to monomer fluorescence ratio of pyrene-PS increased and the fluorescence of NBD-PS was quenched with increasing concentration of PE, it was deduced that PE caused PS-enriched domains in PC/PE/PS membranes. The colocalization of pyrene-PS with BODIPY-PS by PE further supported the possibility. We suggest that PE-induced formation of PS-enriched domains acts as binding sites for apocytochrome c in membranes.

Apoproteins↗
PubMed1999

Phase properties of liquid-crystalline Phosphatidylcholine/Phosphatidylethanolamine bilayers revealed by fluorescent probes.

The mixing properties of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were examined in liquid-crystalline phase using fluorescent probes incorporated into lipid bilayers. The excimer to monomer (E/M) fluorescence ratio of 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine (PPC) versus PPC concentration was higher for binary mixtures containing phosphatidylcholine (PC)/phosphatidylethanolamine (PE) (1:1) compared to PC matrix. When POPC was gradually replaced with POPE, the E/M ratio also increased suggesting the enhanced lateral mobility or the lateral enrichment of PPC into domains or both. Evidences for the PE-induced domain formation were further provided by resonance energy transfer between 2-(4, 4-difluoro-5-methyl-4-boro-3a, 4a-diaza-s-indacene-3-dodecanoyl)-1-hexadecanoyl-sn-glycero- 3-phospho choline and PPC, which was enhanced as a function of PE concentration, and by the polarization of 1,6-diphenyl-1,3, 5-hexatriene. In addition, PE reduced free volume and polarity of lipid bilayers as measured by the emission fluorescence of 1,2-bis PPC and 6-lauroyl-2-dimethylaminonaphthalene. When POPE analogs with a methylated head group instead of normal POPE were used, the diminished effect on the domain formation was shown in the order N-methyl PE > N,N-dimethyl PE. The results suggest that the mixing properties of POPE and POPC are not random but that lipid domains of phospholipids are formed.

2-Naphthylamine↗
PubMed1999

Molecular interactions in ribose transport: the binding protein module symmetrically associates with the homodimeric membrane transporter.

The Escherichia coli high-affinity ribose transporter is composed of the periplasmic ribose-binding protein (RBP or RbsB), the membrane component (RbsC) and the ATP-binding protein (RbsA). In order to dissect the molecular interactions initiating the transport process, RbsC suppressors for transport-defective rbsB mutations were isolated. These suppressors are localized in two regions of RbsC, which are allele-specific to N- or C-terminal domain mutations of RBP, suggesting that there are two distinct regions of RbsC, each interacting with one of the two domains of RBP. To demonstrate that these two regions provide a homodimeric binding surface for RBP we constructed a dimeric rbsC in which two genes are joined tandemly from head to tail with the addition of a linker. The dimeric RbsC protein is stable and functional in growth and ribose uptake. By exploiting the allele specificity between the domain-specific mutations and their suppressors, we generated all mutation-suppressor combinations in a single rbsB plus the dimeric rbsC genes. Their phenotypes are consistent with the proposal that the binding protein module interacts symmetrically with homodimeric RbsC. The mode of association proposed here for the ribose transport components could be extended to other ABC transporters with similar structural organizations.

ATP-Binding Cassette Transporters↗
PubMed1999

Phospholipase D activity of cytochrome P450 in human liver endoplasmic reticulum.

Phospholipase D (PLD) activity in mammalian liver endoplasmic reticulum (ER) has not been characterized. Purified human liver microsomal cytochromes P450 (P450)-P450 1A2 and P450 2E1-were shown to have appreciable PLD activity, hydrolyzing phosphatidylcholine but not other phospholipids, generating PA and choline. The activity was confirmed using recombinant and mutated human P450s expressed in bacteria. In human liver microsomes, immunoinhibition of PLD activity was observed with anti-P450 1A2 > anti-P450 2C > anti-P450 2E1. Thus, P450 may act as a significant PLD in human liver ER and exert its biological effects by several mechanisms, including signaling functions and change of membrane properties.

Amino Acid Sequence↗
PubMed1998

Membrane insertion of cytochrome P450 1A2 promoted by anionic phospholipids.

The role of phospholipids in the membrane binding and subsequent insertion of the microsomal protein rabbit cytochrome P450 (P450) 1A2 into phospholipid bilayers was investigated. The insertion of P450 1A2 into phospholipid bilayers was measured by the quenching of Trp fluorescence of P450 1A2 by pyrene and brominated and doxyl-labeled phospholipids. When the phosphatidylcholine (PC) matrix was replaced with acidic phospholipids [phosphatidic acid (PA), phosphatidylserine, and phosphatidylinositol] and phosphatidylethanolamine (PE), the extent of insertion into lipid bilayers was strictly dependent on the type of acidic phospholipids. All anionic phospholipids caused the penetration of P450 1A2 into lipid bilayers, but PA was the most efficient in facilitating deep penetration of P450 1A2 into bilayers. On the other hand, binding of P450 1A2 to liposomes was increased by acidic phospholipids to the same degree regardless of the type of acidic phospholipids. PE was found to act as an inert matrix phospholipid, similar to PC, as it exerted very little effect on the insertion of P450 1A2 into lipid bilayers and the binding of P450 1A2 to membranes. It was also found that the phospholipid-dependent membrane insertion of P450 1A2 was associated with altered enzyme activity, increased alpha-helix content, and increased Trp fluorescence of P450 1A2. These results indicate that negative charges on the acidic phospholipids are important for the initial binding of P450 1A2 to membranes, but the penetration of P450 1A2 into lipid bilayers is regulated by the type of acidic phospholipids, and that phospholipid-dependent insertion of P450 1A2 is accompanied by a structural change of P450 1A2.

Animals↗
PubMed1998

Effects of nonlamellar-prone lipids on the ATPase activity of SecA bound to model membranes.

The effect of nonlamellar-prone lipids, diacylglycerol (DG) and phosphatidylethanolamine (PE), on the ATPase activity of SecA was examined. When Escherichia coli PE of the standard vesicles composed of 60 mol% of this lipid and 40 mol% of dioleoylphosphatidylglycerol (DOPG) is gradually replaced with either dioleoylglycerol (DOG) or dioeloyl PE (DOPE), the ATPase activity of SecA present together increased appreciably. On the other hand, when E. coli PE of the standard vesicles was replaced with DOG analogs, the SecA ATPase activity decreased slightly, and when replaced with phosphatidylcholine the decrease in the ATPase activity was more appreciable. When DOPE or E. coli PE was added to PC vesicles, the SecA ATPase activity was enhanced only slightly, suggesting that the hexagonal II structure per se is not important for the ATPase activity increase. It was observed that DOG induced phase separation of PG, and the lamellar-hexagonal II (L-HII) transition temperature of vesicles decreased by about 10 degreesC. The DOG analogs had no effect on these properties, suggesting the importance of the phase separation of PG and the decrease of L-HII transition temperature of lipid bilayers to the SecA ATPase activity. The phase separation of PG by Ca2+ also brought about increased ATPase activity of SecA, underlining the importance of phase separation of PG for the enzyme activity. The incorporation of DOG or DOPE in the vesicle also increased the amount of SecA bound to model membranes and the extent of SecA penetration into the membrane. Studies with vesicles without SecA showed increased exposure of hydrophobic acyl chains when the DOG was present. Taken together, these observations suggest that the phase separation of PG and/or the bilayer penetration of SecA are mainly responsible for the enhanced SecA-vesicle interaction with concomitant increase in SecA ATPase activity.

Adenosine Triphosphatases↗
PubMed1998

Conformational change and activation of cytochrome P450 2B1 induced by salt and phospholipid.

A stimulatory effect of increased salt concentration on the enzymatic activity of rat liver microsomes and a reconstituted system containing cytochrome P450 (P450) 2B1 and NADPH-P450 reductase was seen. Structural change of P450 2B1 accompanying the salt-induced increase in its enzyme activity was investigated by circular dichroism, fluorescence spectroscopy, and absorption spectroscopy. It was found that the salt increased alpha-helix content of P450 2B1 in the presence as well as in the absence of a phospholipid. Intrinsic fluorescence emissions also increased with increasing salt concentration. The low-spin iron configuration of P450 2B1 shifted toward the high-spin configuration in response to the increased salt concentration. It was found that the activity increase of P450 coincides with the raised alpha-helix content. The presence of phospholipid magnified this effect. It is proposed that the interaction with salts and phospholipid molecules surrounding P450 2B1 in the endoplasmic reticulum is important for a functional conformation of P450 2B1 in a monooxygenase system including NADPH-P450 reductase.

Animals↗
PubMed1998

Phase separation in phosphatidylcholine/anionic phospholipid membranes in the liquid-crystalline state revealed with fluorescent probes.

The mixing properties of anionic phospholipids such as phosphatidic acid (PA), phosphatidylserine (PS), and phosphatidylglycerol (PG) with phosphatidylcholine (PC) were examined in the liquid-crystalline state of membranes using extrinsic fluorescent probes incorporated into lipid bilayers. The excimer to monomer (E/M) fluorescence ratio of 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine (PPC) was higher for the PA and PS matrices as compared to that for the PC matrix. When PC was replaced with PA or PS, the E/M ratio of PPC also increased in a concentration-dependent manner. When the concentration of PA or PS was increased in the PC membrane, the fluorescence of 1-palmitoyl-2-[12-[7-nitro-2,1, 3-benzodiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine decreased, indicating the occurrence of lipid clustering. Direct evidence for the PA or PS-induced phase separation in the PC/PA or PC/PS system was provided by the resonance energy transfer between 2-(4,4-difluoro-5-methyl-4-boro-3a, 4a-diaza-s-indacene-3-dodecanoly)-1-hexadecanoly -sn-glycero-3-phospho choline and PPC. The fluorescence polarization of 1,6-diphenyl-1,3, 5-hexatriene further supported the lateral organization of membranes by PA and PS. PA and PS also reduced the polarity of lipid bilayers, as measured by the emission fluorescence of 6-lauroyl-2-dimethylaminonaphthalene. On the other hand, PG had very little effect on the PC matrix, suggesting the ideal miscibility with PC molecules. The results suggest that the mixing properties of PA and PS in the PC matrix are not random but that phase separation occurs in the liquid-crystalline phase of membranes.

Anions↗
PubMed1998

[Diagnostic usefulness of magnetic resonance cholangiopancreatography (MRCP) in comparison with retrograde cholangiopancreatography (ERCP) for cholelithiasis].

PURPOSE: To elucidate MRCP diagnostic usefulness compared to ERCP. PATIENTS, METHODS: 29 MRCP performed patients diagnosed by ERCP were studied. RESULTS: MRCP showed 92%, 67% sensitivity and 73%, 100% specificity in choledocholithiasis (CBD) and cholecystolithiasis (GB), respectively. MRCP revealed 4.0 false positive and 1.5 false negative in CBD and GB, respectively. MRCP could detect 22 stones out of 33 in CBD and 13 out of 42 in GB. As for the maximal diameter, MRCP depicted 31% for less than 5 mm and 100% for over 6 mm in CBD. In GB, 7% for less than 5 mm, 83% for 6-10 mm and 100% for over 11 mm. CONCLUSION: Although the depection of tiny stone is limited, MRCP is a useful diagnostic tool for cholelithiasis, especially choledocholithiasis.

Adult↗
PubMed1997

Hepatosplenic T-cell lymphoma: prolymphocytic transformation 18 months after splenectomy.

We report a young woman with pancytopenia and huge splenomegaly who was also found to have peripheral T-cell lymphoma with massive infiltration of T-cell evident in the liver and spleen. A liver biopsy showed predominant sinusoidal infiltration of pan-T cell antibody-stained T-lymphoid cells. Histologic examination of the spleen revealed numerous tumor cells predominantly infiltrated in the cords and sinuses of the red pulp, which were identical to those described in the liver. Several clusters of small round abnormal cells were observed in marrow cytology. Although the patient felt well during 18 months after the splenectomy was done, the patient eventually manifested a huge hepatomegaly, showed increasing white blood cell count to 42 x 10(9)/l, and numerous prolymphocytes (66.9%) in the bone marrow. This change represented a prolymphocytic transformation of the patient's original hepatosplenic T-cell lymphoma.

Adult↗
PubMed1996

Conformational change of cytochrome P450 1A2 induced by sodium chloride.

Recently, it was reported that the activity of rabbit P450 1A2 is markedly increased at elevated sodium phosphate concentration. Here, the possible structural change of rabbit P450 1A2 accompanying the NaCl-induced increase in its enzyme activity is investigated by fluorescence spectroscopy, circular dichroism, and absorption spectroscopy. It was found that NaCl increased alpha-helix content and lowered beta-sheet content of P450 1A2 in the presence as well as in the absence of a phospholipid. Intrinsic fluorescence emissions also increased with increasing NaCl concentration. The low spin iron configuration of P450 1A2 shifted toward the high spin configuration in response to the increased salt concentration. The effect of increased potassium phosphate and NaCl on the P450 1A2 activity was also studied. It was found that the activity increase of rabbit P450 1A2 occurs concomitantly with the conformational change including raised alpha-helix content.

Animals↗