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- きみとし かやぬま
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1 -DLZnsodium zinc dihydrolipoylhistidinatezn2+/ -- B-16 -α PLC- Lineweaver-Burk - -
2 (1/3) 2
3 (2/3) ~~ - + R=/C R 2 2,, C
4 (3/3) ~~ 4 DPAchrome (µm) DPA only + DLA + LA α-ladla DLA DLA Time (min) xidation by Tyrosinase DLA DPA DPAquinone 5--LipoylDPA * Tsuji-aito, K.; atani, T.; kada, T.; Tehara, T. Biochem. Biophys. Res. Commun. 26, 343, 15.
5 - -DLZnsodium zinc dihydrolipoylhistidinate Zn 2+ / Zn a - --
6 -B16 75M % DLAα- 36.3%38.3% DLA DLZn α-arbutin control 1 5 -B16 - α- Inhibitory effect of melanin synthesis (%)
7 1 5 - LADLA * - -DLA α-dla - LA DLA DLZn -Cystein L α-arbutin Kojic acid * Tsuji-aito, K.; atani, T.; kada, T.; Tehara, T. Biochem. Biophys. Res. Commun. 26, 343, 15. control Inhibitory effect of tyrosinase (%)
8 -- (1/4) DLA a -
9 -- (2/4) Absorbance at 26 nm Relative Intensity (%) 1 5 Peak II Peak I Peak III Time (min) m/z Wavelength (nm) [M+] m/z The three molecular ion peaks of the expected monoconjugate product, named peak I, II (a shoulder peak of peak III), and III, were detected at m/z 541; the substances coeluted at the time characteristic of the major reaction products. Absorbance (mau) -- UV EI+-MPLC m/z 5413 IIIIII III DLA 256nm291nm 3DLA
10 -- (3/4) III PLCII Relative Intensity (%) C. a mw m/z [M+] M/M M/M -DLA m/z DLA
11 -- (4/4) 1 A. - DLZn 5 M/M B. + DLZn - - -B16 Relative Intensity (%) M/M C. + DLZn M/M PLC/M/M m/z 541. [M + ] B D. Reserpine (Internal standard) M/M Time (min)
12 - (1/2) DL DPA conjugate(s) / reserpine (rate of peak area counts) Peak II + III Peak I Incubaton time (hour) - m/z 69. [M + ] PLC/M/M - B DL DPA conjugate(s) / reserpine (rate of peak area counts) Peak II + III Peak I Peak II + III Peak I DPA concentration (µm) DLZn concentration (µm)
13 - (2/2) Absorbance at 475 nm control DLZn Incubation time (hour)
14 - (1/2) - - DLZn IC5 (µm) 1.33 K m (M) 4.7 x 1-3 V m ( A 492 /min) 1.28 x 1-1 K I (M) 3.56 x 1-7 Inhibitor type competitive DLZn. µm.5 µm 1. µm 1.5 µm 2. µm 1/v ( A 492 /min) /[] (mm -1 ) - 4.µMIC51.3µM82.9% - - Km Vmax Vmax DA492/min1.3 x 1-4Km.86mM 1.3µM- Vmax1.3 x 1-4 Km4.1 x DPA - -
15 - (2/2) E + E E + P 1 [ 2 ] P 1 P 2 Lineweaver-Burk- P' 1 [DLZn] E: tyrosinase : L -DPA P 1 : DPAquinone P 2 : DPAchrome (Abs 492 nm) P 1 : DL DPA conjugated product(s) - - -
16 DPA-quinone Tyrosinase Tyrosine Tyrosinase DPA-quinone - DLZn - + DPA-chrome R = /C DL DPA conjugate a 2 Uncharacterized series of reactions Cysteine Cysteinyl-DPA Indoles Benzothiazine 2 R Eumelanin Modulation of melanin synthesis Pheomelanin
17 -DLZnsodium zinc dihydrolipoylhistidinate
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