Continuous pH-stat titration method for the assay of lipoprotein lipase activity in vitro
A method was described where the hydrolysis of triolein to fatty acids and glycerol by lipoprotein lipase (LpL) was continuously monitored by titration of the liberated protons in a pH-stat automatic titrator. The method gave results comparable to those obtained by a conventional assay, and was also found to be applicable to the study of crude and partially purified enzyme preparations from both cow milk and rat adipose tissue. In agreement with earlier results obtained by other methods, the first product of hydrolysis was identified as 1,2-(2,3-)diglyceride, which was further converted to 2- or 1-monoglyceride. A good comparison between the titration and conventional methods was also obtained when peptides from human serum very low density lipoproteins (VLDL) were tested either as activators or inhibitors of LpL. Thus, the method appears suitable also for a systematic investigation of the role of substrates, activator(s) and inhibitors on LpL activity in vitro.
- Research Organization:
- Univ. of Chicago
- OSTI ID:
- 5263422
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 62; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ALCOHOLS
ANIMALS
BIOASSAY
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BODY FLUIDS
CARBOXYLIC ACIDS
ENZYMES
ESTERASES
FOOD
GLYCEROL
HYDROLASES
HYDROXY COMPOUNDS
IN VITRO
KINETICS
LIPASES
LIPIDS
LIPOPROTEINS
MAMMALS
MILK
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
RATS
REACTION KINETICS
RODENTS
VERTEBRATES