Substrate specificity of the isoprenylated protein endoprotease.

Substrate specificity of the isoprenylated protein endoprotease.
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异戊二烯化蛋白质内切蛋白酶的底物特异性。

DOI:
10.1021/bi00162a014
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发表时间:
1992
期刊:
影响因子:
2.9
通讯作者:
Rando,RR
Rando,RR
中科院分区:
生物学3区
文献类型:
--
作者:
Ma,YT;Chaudhuri,A;Rando,RR

文献摘要

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方法类似物的合成。Cys-Val-Ile和ZerZ-丁硫基-Cys-Val-Ile是由Chuck Dahl博士在哈佛医学院多肽合成设施中合成的。异戊二烯基化肽类似物的制备方法与已发表的方法相似或相同(Ma&Rando,1992)。合成方案的完整细节作为补充材料提供。核磁共振数据和类似物的层析行为如下所述。N-[3H\Acetyl-S-ali-trans-farnesyl-L-Cys-D-Val-L-Ile.采用高效液相色谱法(Rainin Silica,4.6×250 mm,正己烷/异丙醇/TFA(92:8:0.01)为洗脱剂,流速1.5mL/min,检测波长210 nm)对多肽进行纯化。低密度脂蛋白异构体的保留时间为8.60min:核磁共振(DMSO-t/e,500 MHz),8.11(1H,d,J=9.5 Hz),8.09(1H,d,J=7.5 Hz),8.01(1H,d,i=7.5 Hz),5.15(1H,t,i=8 Hz),5.04(2H,t,i=8 Hz),4.59(1H,dd,J=8,15 Hz),4.34(1H,dd,i=8,15 Hz),=6,9赫兹),4.18(1 H,dd,J=6.5,8.5赫兹),3.17(2 H,dd,=4,7.5赫兹),2.65(1 H,dd,J=6.5,13.5赫兹),2.51(1 H,dd,J=7.5,13.5赫兹),2.08-1.90(9 H,m),1.80(3 H,S),1.79(1 H,m),1.61(3 H,S),1·60(3 H,S),1·54(6 H,S),1·38(1 H,m),1·19(1 H,m),0·79-0·87(12 H,m)。N-[~3H]Acetyl-S-all-trans-farnesyl-L-Cys-L-Val-D-Ile.LLD异构体的保留时间为8.19分钟:核磁共振(DMSO-
MethodsSyntheses of Analogs. Cys-Val-Ile and ZerZ-butylthio-Cys-Val-Ile were synthesized by Dr. Chuck Dahl at the Harvard Medical School peptide synthesis facility. The isoprenylated peptide analogs were prepared by methods similar or identical to those publishedearlier (Ma & Rando, 1992). Complete details of the synthetic protocols are provided as supplementary material. The NMR data and chromatographic behavior of the analogs are described below. N-[3H\Acetyl-S-ali-trans-farnesyl-L-Cys-D-Val-L-Ile. The peptide was purified by HPLC (Rainin silica, 4.6 X 250 mm, hexane/2-propanol/TFA (92: 8: 0.01) as eluant at a flow rate of 1.5 mL/min with UV detection at 210 nm). The retention time of the ldl isomer was 8.60 min: NMR (DMSO-t/e, 500 MHz), 8.11 (1 H, d, J= 9.5 Hz), 8.09 (1 H, d, J= 7.5 Hz), 8.01 (1 H, d,/= 7.5 Hz), 5.15 (1 H, t,/= 8 Hz), 5.04 (2 H, t,/= 8 Hz), 4.59 (1 H, dd, J= 8, 15 Hz), 4.34 (1 H, dd,/=6, 9 Hz), 4.18 (1 H, dd, J= 6.5, 8.5 Hz), 3.17 (2 H, dd,/= 4, 7.5 Hz), 2.65 (1 H, dd, J= 6.5, 13.5 Hz), 2.51 (1 H, dd, J= 7.5, 13.5 Hz), 2.08-1.90 (9 H, m), 1.80 (3 H, s), 1.79 (1 H, m), 1.61 (3 H, s), 1.60 (3 H, s), 1.54 (6 H, s), 1.38 (1 H, m), 1.19 (1 H, m), 0.79-0.87 (12 H, m). N-[3H] Acetyl-S-all-trans-farnesyl-L-Cys-L-Val-D-Ile. The retention time of the lld isomer was 8.19 min: NMR (DMSO-