Stopped-Flow Analysis on Anion Binding to Blue-Form Halorhodopsin fromNatronobacterium pharaonis:  Comparison with the Anion-Uptake Process during the Photocycle†

Biochemistry ◽  
2002 ◽  
Vol 41 (7) ◽  
pp. 2452-2458 ◽  
Author(s):  
Maki Sato ◽  
Tatsuaki Kanamori ◽  
Naoki Kamo ◽  
Makoto Demura ◽  
Katsutoshi Nitta
Keyword(s):  
Flow Analysis ◽  
Anion Binding ◽  
Stopped Flow ◽  
Uptake Process ◽  
Anion Uptake

Stopped-flow analysis on the mechanism of perylene nanoparticle formation by the reprecipitation method

2009 ◽  
Vol 311 (3) ◽  
pp. 553-555 ◽  
Author(s):  
Junichi Mori ◽  
Yousuke Miyashita ◽  
Daniel Oliveira ◽  
Hitoshi Kasai ◽  
Hidetoshi Oikawa ◽  
...  
Keyword(s):  
Flow Analysis ◽  
Stopped Flow ◽  
Nanoparticle Formation ◽  
Reprecipitation Method

Human RhAG ammonia channel is impaired by the Phe65Ser mutation in overhydrated stomatocytic red cells

2012 ◽  
Vol 302 (2) ◽  
pp. C419-C428 ◽  
Author(s):  
Sandrine Genetet ◽  
Pierre Ripoche ◽  
Julien Picot ◽  
Sylvain Bigot ◽  
Jean Delaunay ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Hemolytic Anemia ◽  
Rate Constant ◽  
Rate Constants ◽  
Flow Analysis ◽  
Red Cells ◽  
Stopped Flow ◽  
Loss Of Function ◽  
Dramatic Decrease ◽  
Time Courses

In red cells, Rh-associated glycoprotein (RhAG) acts as an ammonia channel, as demonstrated by stopped-flow analysis of ghost intracellular pH (pHi) changes. Recently, overhydrated hereditary stomatocytosis (OHSt), a rare dominantly inherited hemolytic anemia, was found to be associated with a mutation (Phe65Ser or Ile61Arg) in RHAG. Ghosts from the erythrocytes of four of the OHSt patients with a Phe65Ser mutation were resealed with a pH-sensitive probe and submitted to ammonium gradients. Alkalinization rate constants, reflecting NH3transport through the channel and NH3diffusion unmediated by RhAG, were deduced from time courses of fluorescence changes. After subtraction of the constant value found for Rhnulllacking RhAG, we observed that alkalinization rate constant values decreased ∼50% in OHSt compared with those of controls. Similar RhAG expression levels were found in control and OHSt. Since half of the expressed RhAG in OHSt most probably corresponds to the mutated form of RhAG, as expected from the OHSt heterozygous status, this dramatic decrease can be therefore related to the loss of function of the Phe65Ser-mutated RhAG monomer.

scholarly journals 1PT164 Functional and Stopped-flow analysis of circularly permutated GroEL mutant CP86(The 50th Annual Meeting of the Biophysical Society of Japan)

2012 ◽  
Vol 52 (supplement) ◽  
pp. S97
Author(s):  
Toshifumi Mizuta ◽  
Tatsuya Uemura ◽  
Takuma Hirayama ◽  
Kunihiro Hongo ◽  
Yasushi Kawata ◽  
...  
Keyword(s):  
Annual Meeting ◽  
Flow Analysis ◽  
Stopped Flow ◽  
Biophysical Society

Role of RhAG and AQP1 in NH3 and CO2 gas transport in red cell ghosts: a stopped-flow analysis

2006 ◽  
Vol 13 (1-2) ◽  
pp. 117-122 ◽  
Author(s):  
P. Ripoche ◽  
D. Goossens ◽  
O. Devuyst ◽  
P. Gane ◽  
Y. Colin ◽  
...  
Keyword(s):  
Gas Transport ◽  
Flow Analysis ◽  
Stopped Flow ◽  
Red Cell ◽  
Co2 Gas

scholarly journals Proton release on binding of glutathione to Alpha, Mu and Delta class glutathione transferases

1999 ◽  
Vol 344 (2) ◽  
pp. 419-425 ◽  
Author(s):  
Anna Maria CACCURI ◽  
Giovanni ANTONINI ◽  
Philip G. BOARD ◽  
Michael W. PARKER ◽  
Maria NICOTRA ◽  
...  
Keyword(s):  
Flow Analysis ◽  
Three Dimensional ◽  
Kinetic Mechanism ◽  
Activation Mechanism ◽  
Dimensional Structure ◽  
Binary Complex ◽  
Stopped Flow ◽  
Binding Mechanism ◽  
Proton Extrusion ◽  
Evolutionary Pathway

Potentiometric, spectroscopic and stopped-flow experiments have been performed to dissect the binding mechanism of GSH to selected glutathione S-transferases (GSTs), A1-1, M2-2 and Lucilia cuprina GST, belonging to Alpha, Mu and Delta classes respectively. Both Alpha and Mu isoenzymes quantitatively release the thiol proton of the substrate when the binary complex is formed. Proton extrusion, quenching of intrinsic fluorescence and thiolate formation, diagnostic of different steps along the binding pathway, have been monitored by stopped-flow analysis. Kinetic data are consistent with a multi-step binding mechanism: the substrate is initially bound to form an un-ionized pre-complex [k1⩾ (2-5)×106 M-1˙s-1], which is slowly converted into the final Michaelis complex (k2 = 1100-1200 s-1). Ionization of GSH, fluorescence quenching and proton extrusion are fast events that occur either synchronously or rapidly after the final complex formation. The Delta isoenzyme shows an interesting difference: proton extrusion is almost stoichiometric with thiolate formed at the active site only up to pH 7.0. Above this pH, at least one protein residue acts as internal base to neutralize the thiol proton. These results suggest that the Alpha and Mu enzymes retain not only a similar catalytic outcome and overall three-dimensional structure but also share a similar kinetic mechanism for GSH binding. The Delta GST, which is closely related to the mammalian Theta class enzymes and is distantly related to Alpha and Mu GSTs in the evolutionary pathway, might display a different activation mechanism for GSH.

Time-resolved chemiluminescence: a novel tool for improved emission sequence in stopped-flow analysis

2004 ◽  
Vol 517 (1-2) ◽  
pp. 111-117 ◽  
Author(s):  
José A Murillo Pulgarı́n ◽  
Luisa F Garcı́a Bermejo ◽  
José A Rubio Aranda
Keyword(s):  
Flow Analysis ◽  
Stopped Flow ◽  
Time Resolved

Stopped-Flow Analysis of CO and NO Binding to Inducible Nitric Oxide Synthase†

Biochemistry ◽  
1998 ◽  
Vol 37 (11) ◽  
pp. 3777-3786 ◽  
Author(s):  
Husam M. Abu-Soud ◽  
Chaoqun Wu ◽  
Dipak K. Ghosh ◽  
Dennis J. Stuehr
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Flow Analysis ◽  
Stopped Flow ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Methionyl-tRNA synthetase from Escherichia coli: active stoichiometry and stopped-flow analysis of methionyl adenylate formation

Biochemistry ◽  
1976 ◽  
Vol 15 (17) ◽  
pp. 3678-3685 ◽  
Author(s):  
Francois Hyafil ◽  
Yannick Jacques ◽  
Guy Fayat ◽  
Michel Fromant ◽  
Philippe Dessen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Flow Analysis ◽  
Trna Synthetase ◽  
Stopped Flow ◽  
Methionyl Trna Synthetase
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