(Quality of Life; QL) QL (Nerve Growth Factor; NGF) NGF PC12 6- (6-HITC)
NGF NGF NGF PTP1B 6-HITC PTP1B NGF PTP1B 6-HITC PTP1B 6-HITC NGF 6-HITC 6-HITC in vivo 464-8601 Tel: 052-789-4126
20-Hydroxyecdysone Dibenzoylhydrazine
ABA 1960 ABA ABA ABA ABA ABA ABA ABA ABA ABA ABA ABA 8'- P450 ABA ABA
ABA ABA ABA 8'- 422-8529 836 TEL&FAX054-238-4871 E-mailw5613005@ipc.shizuoka.ac.jp
NH PP N R H H HH NH H P P N H H HH R UDP H AcHN H H H H H H H H C - H H AcHN H P P - - AcHN H H H P - NH N HH P - NH N HH UDP-LacNAc Kobata, J. Biochem., 53, 167 (1963). UDP-Neu5AcLacNAc Jourdian et al., Fed. Proc., 20, 161 (1961). UDP E-mail : j6103006@guedu.cc.gifu-u.ac.jp Tel : 058-293-2916
Flammulina velutipes F. velutipes PCR fluorescence differential display (FDD) 10 RNA FDD cdna 75 46 accession No. AB201060-AB201106 17 northern blot BLASTX 29 cdna GTP-binding protein growth factor ubiqutin-proteasome cytochrome P450 hydrophobin polysaccharide deacetylase
Fluorescence Differential Disply
Wild-type rsp5 mutant YPD 25 YPD 37 SD 37 Ubiquitination Ub Ub Growth No Ubiquitination Growth inhibition Rsp5 Ub Ub Ub Abnormal proteins Degradation in proteasome or vacuole rsp5 Ub Abnormal proteins accumulation Wild-type rsp5 mutant + Temp. up-shift + Sorbitol + Ethanol Wild-type 0 30 60 120 0 30 60 120 Time min HSP12 ACT1 rsp5 mutant Time min Wild-type rsp5 mutant V H M V H -HA western -Pgk1 western M Hsf1 Msn4 Pgk1 Golgi ER proteasome Ub Ub Rsp5 Ubiquitination? Hsf1 HSE Ub Hsf1 Msn2/4 STRE Ub Msn2/4 Transport? Nucleus Induction HSP42, HSP12, DDR2
Figure 1 H 3 S H NH2 R Me N H N NH H 2 C N Me H H 1a: Liposidomycin A : R= Me (CH 2 ) 3 1b: Liposidomycin B : R= 1c: Liposidomycin C : R= CH 2 Me 2 (CH 2 ) 8 Me(CH 2 ) 10 H H 2 C N Me Me N H H N H degradation product (2) NH
Figure 2 Me H N H N H 2 C N Me H H 2 NH Me Bz N Bn N Ph N Me Ac Ac 3 NH Ph NH Me 4 + H N 3 5 Bn Me H Bz H Ph N Me H D-ribose Me N 7 H Bn 5 Me (CCl) 2, DIPEA CH 2 Cl 2 4 (11steps 20% from D-ribose) 2 steps 41% Bz Ph Me Ph N Me N N 3 6 Ac 8 Bn Me N Bn Ac Ac Me 7steps 11% TMS N N SnCl 4 9 TMS 3 Figure 3 Bn Ac N Bn 4 steps 9,SnCl 4 N NH N 37% Ns 55% R Ac Ac Ac Ac Bn N Me 10 11: R=Ns 13 12: R=CH Bz R R N Bn N Bn Bz Ac N NH Ph N Ph N Me Ac Ac Me Ac Ac 14 R=Ns or CH 15 N NH 3 2
( TGase TGase 8 TGase 1, 3 Factor XIII TGase 2 TGase TGase TGase M13 piii N 12 TGase
TGase TGase 2 Factor XIII TGase N
- (Caco-2 ) (HepG2 ) I-I-A-E-K ( ) Ile Ile Ala Glu Lys - - - - Biochem. Biophys. Res. Commun. 281, 11-17 (2001)
(RAS) (ACE) RAS D R V Y I H P F H L V D R V Y I H P F H L D R V Y I H P F RAS 10 ACE Na His P5 -Pro P4 -Phe P3 (HPF)
1) HPF HPF (VA) WSZ1a BSZ4 8 RADHPF( III) ADHPFLFLIK(VA-X ) ADHPFLFFIR(VA-Y ) DHPFLFLV HPFLFLVR(WSZ1a ) VANHPFLFLIRVANHPFLFLIR(BSZ4 ) (TRX)C (TRX- ) BL21(DE3) TRX TRX (300 µm) TRX 100 TRX-ADHPFLFLIKTRX-ADHPFLFFIR TRX-DHPFLFLVTRX-HPFLFLVRTRX-VANHPFLFLIR TRX-ADHPFLFLIKTRX-ADHPFLFFIRTRX-DHPFLFLVTRX-HPFLFLVR TRX-VANHPFLFLIR(300 µm) 25 28 57 62 58 IC50 600600 2707580 µm TRX-VANHPFLFLIR Ki 53 µm TRX-RADHPFTRX-VANHPFTRX-LFLIR(300 µm) HPF C () () 1) 2002 p. 123
γ [ ] 3 ATP 2 γ GCS γ 2 γ GCS
GCS buthionine sulfoximine(bs) BS GCS GCS BS GCS BS X [ ] GCSBSMgADP PEG10,000 X SPring-8 BL40B2 2.3Å P21 a=70.5 Å, b=97.6 Å, c=102.7 Å, β=109.5 R 21%(Rfree24) GCSBSMgADP ATP γ BS BS TSA GCS α TSA Tyr241 Tyr300 GCS GCSTSA BS Tyr300 Tyr300 BS TSA 500 Tyr300 Tyr241 Tyr241 Tyr241 BS CH-π Tyr241 Gln144