Severe MgADP Inhibition of <i>Bacillus subtilis</i> F₁-ATPase Is Not Due to the Absence of Nucleotide Binding to the Noncatalytic Nucleotide Binding Sites

<div><p>F₁-ATPase from <i>Bacillus subtilis</i> (BF₁) is severely suppressed by the MgADP inhibition. Here, we have tested if this is due to the loss of nucleotide binding to the noncatalytic site that is required for the activation. Measurements with a tryptophan mutant of BF₁ indicated that the noncatalytic sites could bind ATP normally. Furthermore, the mutant BF₁ that cannot bind ATP to the noncatalytic sites showed much lower ATPase activity. It was concluded that the cause of strong MgADP inhibition of BF₁ is not the weak nucleotide binding to the noncatalytic sites but the other steps required for the activation.</p></div

Emission spectra of α₃β₃γ^WT and α₃β₃γ^RW complexes of BF₁.

<p>Fluorescence emission spectra of α₃β₃γ^WT and α₃β₃γ^RW complexes in the absence and presence of 1 mM ATP are shown. Excitation wavelength was 300 nm and the fluorescence emission spectra were recorded at 50 nm/min. Excitation and emission slit-widths were set at 5 and 10 nm, respectively. The fluorescence values are normalized to peak of α₃β₃γ^RW in the absence of ATP as 100%. Solid line, dotted line, two-dot-chain line and dashed line represent α₃β₃γ^RW-ATP, α₃β₃γ^RW+ATP, α₃β₃γ^WT-ATP and α₃β₃γ^WT+ATP, respectively.</p

Titration of fluorescence by ATP.

<p>Changes in the fluorescence upon addition of ATP are expressed as percent of the fluorescence before the addition of ATP. Values are taken when the fluorescence reached a plateau. Error bars represent standard errors. The solid line represents the theoretical curve with the Hill equation (Fluorescence quenching  = <i>ΔFL</i>_max×[ATP]ⁿ/(<i>K_d</i>ⁿ+[ATP]ⁿ)) with the following parameters(± standard error); <i>K_d</i> = 36.5±1.2 µM, <i>ΔFL</i>_max = 71.0±0.7%, <i>n</i> = 1.47±0.1. The dotted line represents the theoretical curve with simple binding equation (Fluorescence quenching  = <i>ΔFL</i>_max×[ATP]/(<i>K_d</i>+[ATP])) with <i>K</i>_d = 34.4±2.9 µM, <i>ΔFL</i>_max = 73.5±1.5%.</p

ATPase activities of α₃β₃γ complexes.

<p>The initial (A) and steady-state (B) ATPase activities and ATPase activity in the presence of 0.1% LDAO (C) were determined. Hatched, open and solid bars represent α₃β₃γ^WT, α₃β₃γ^RW and α₃β₃γ^ΔNC, respectively. Error bars represent standard errors. Asterisks represent statistically significant differences (*p<0.05 and **p<0.01, Student's t-test). Data for WT are taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107197#pone.0107197-Mizumoto1" target="_blank">[12]</a>.</p

Comparison of ROI on conventional CT (scan parameter; 120kVp, 1366mAs, total scan time; 2.8 seconds, slice thickness; 0.98mm) and CBCT (scan parameter; 119kVp, 165mA, total sweep time; 5.2 seconds, slice thickness; 0.68mm)

<p>Comparison of ROI on conventional CT (scan parameter; 120kVp, 1366mAs, total scan time; 2.8 seconds, slice thickness; 0.98mm) and CBCT (scan parameter; 119kVp, 165mA, total sweep time; 5.2 seconds, slice thickness; 0.68mm)</p

a) Correlation of CB-CT and conventional CT values. b) Correlation of CB-CT and conventional CT values. c) Correlation of CB-CT and conventional CT values.

<p>a) Correlation of CB-CT and conventional CT values. b) Correlation of CB-CT and conventional CT values. c) Correlation of CB-CT and conventional CT values.</p

Local recurrences after balloon occuluded transcatheter arterial chemoembolization with miriplatin.

<p>The local recurrence rates at 6 months, and 1 year were 11.1, and 26.2%, respectively.</p

Clinical background of 51 patients.

<p>SD; Standard Deviation, AFP; alpha-fetoprotein, DCP; des-gamma-carboxy prothrombin.</p

In regard to plain CT results immediately after B-TACE treatment, the local recurrence rate in the higher-than-mean CT value group was 4.8% at 6 months and 16.0% at 12 months, but in the lower-than-mean CT value group, the local recurrence rates were significantly different: 15.2% at 6 months and 32.9% at 12 months.

<p>In regard to plain CT results immediately after B-TACE treatment, the local recurrence rate in the higher-than-mean CT value group was 4.8% at 6 months and 16.0% at 12 months, but in the lower-than-mean CT value group, the local recurrence rates were significantly different: 15.2% at 6 months and 32.9% at 12 months.</p

Risk factors for local recurrence after MPT-B-TACE.

<p>HR; Hazard Ratio, CI; Confidence Interval, AFP; alpha-fetoprotein, DCP; des-gamma-carboxy prothrombin.</p