Measurement of diacylglycerol acyltransferase activity in isolated hepatocytes

被引：4
|
作者：
Geelen, MJH [1 ]
机构：
[1] Univ Utrecht, Fac Vet Med, Dept Biochem & Cell Biol, NL-3508 TD Utrecht, Netherlands
来源：
ANALYTICAL BIOCHEMISTRY | 2003年 / 322卷 / 02期
关键词：
triacylglycerol synthesis; diacylglycerol acyltransferase; rat hepatocytes; digitonin;
D O I：
10.1016/j.ab.2003.08.019
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
An assay procedure for diacylglycerol acyltransferase that allows rapid measurement of the activity of this enzyme in isolated hepatocytes is described. The one-step procedure involves permeabilization of the plasma membrane with digitonin and simultaneous measurement of diacylglycerol acyltransferase activity. Digitonin at a concentration of 64 mug/mg of cellular protein was found to be optimal for exposing microsomal diacylglycerol acyltransferase to the components of the assay. The enzyme assay is linear with time up to 4 min and with protein concentrations in the range 0.25-2.4 mg of cellular protein/assay. It is shown that there is a good correlation of cellular enzyme activity as determined in digitonin-permeabilized hepatocytes with the rate of triacylglycerol synthesis in intact hepatocytes. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码：264 / 268
页数：5
相关论文
共 50 条
  • [21] Inhibitory activity of diacylglycerol acyltransferase by tanshinones from the root of Salvia miltiorrhiza
    Ko, JS
    Ryu, SY
    Kim, YS
    Chung, MY
    Kang, JS
    Rho, MC
    Lee, HS
    Kim, YK
    ARCHIVES OF PHARMACAL RESEARCH, 2002, 25 (04) : 446 - 448
  • [22] REGULATION OF TRIACYLGLYCEROL SYNTHESIS IN THE LIVER - MODULATION OF DIACYLGLYCEROL ACYLTRANSFERASE ACTIVITY INVITRO
    HAAGSMAN, HP
    DEHAAS, CGM
    GEELEN, MJH
    VANGOLDE, LMG
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1982, 257 (18) : 593 - 598
  • [23] Inhibitory activity of diacylglycerol acyltransferase by tanshinones from the root ofSalvia miltiorrhiza
    Jeong Suk Ko
    Shi Young Ryu
    Young Sup Kim
    Mi Yeon Chung
    Jong Seong Kang
    Mun-Chual Rho
    Hyun Sun Lee
    Young Kook Kim
    Archives of Pharmacal Research, 2002, 25 : 446 - 448
  • [24] Inhibition of Diacylglycerol Acyltransferase 2 Versus Diacylglycerol Acyltransferase 1: Potential Therapeutic Implications of Pharmacology
    Amin, Neeta B.
    Saxena, Aditi R.
    Somayaji, Veena
    Dullea, Robert
    CLINICAL THERAPEUTICS, 2023, 45 (01) : 55 - 70
  • [25] Diacylglycerol acyltransferase activity and triacylglycerol synthesis in germinating castor seed cotyledons
    He, XH
    Chen, GQ
    Lin, JT
    McKeon, TA
    LIPIDS, 2006, 41 (03) : 281 - 285
  • [26] Enhanced antitumor activity of xanthohumol, a diacylglycerol acyltransferase inhibitor, under hypoxia
    Goto, K
    Asai, T
    Hara, S
    Namatame, I
    Tomoda, H
    Ikemoto, M
    Oku, N
    CANCER LETTERS, 2005, 219 (02) : 215 - 222
  • [27] Abietane diterpenoids of Rosmarinus officinalis and their diacylglycerol acyltransferase-inhibitory activity
    Cui, Long
    Kim, Mun Ock
    Seo, Jee Hee
    Kim, Il Soon
    Kim, Nam Ye
    Lee, Sun Hwa
    Park, Jeongjun
    Kim, Jungwoo
    Lee, Hyun Sun
    FOOD CHEMISTRY, 2012, 132 (04) : 1775 - 1780
  • [28] CHARACTERIZATION OF THE DIACYLGLYCEROL ACYLTRANSFERASE ACTIVITY IN THE MEMBRANE-FRACTION FROM A FUNGUS
    KAMISAKA, Y
    YOKOCHI, T
    NAKAHARA, T
    SUZUKI, O
    LIPIDS, 1993, 28 (07) : 583 - 587
  • [29] REGULATION OF TRIACYLGLYCEROL SYNTHESIS IN PERMEABILIZED RAT HEPATOCYTES - ROLE OF FATTY-ACID CONCENTRATION AND DIACYLGLYCEROL ACYLTRANSFERASE
    STALS, HK
    TOP, W
    DECLERCQ, PE
    FEBS LETTERS, 1994, 343 (01) : 99 - 102
  • [30] Inhibition of diacylglycerol acyltransferase by prenylated flavonoids isolated from the stem bark of Maackia amurensis
    Li, Na
    Zhang, Nan
    Li, Jia-Lin
    Du, Pei-Ge
    Lee, Hyun-Sun
    Cui, Long
    JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH, 2015, 17 (02) : 178 - 181
12345