Degree Type
Dissertation
Date of Award
1986
Degree Name
Doctor of Philosophy
Department
Biochemistry, Biophysics and Molecular Biology
Abstract
Hydrolysis of vitamin A esters by liver is important during their uptake as part of chylomicron remnants and for their mobilization from storage in vitamin A-containing globules. The regulation and properties of the enzyme thought to catalyze these reactions, retinyl ester hydrolase (REH), were studied by the use of a sensitive in vitro assay developed for this enzyme in pig and rat liver. Hydrolysis of retinyl palmitate was maximal in 3- (3-cholamidopropyl) dimethylammonio -1-propanesulfonate (CHAPS) and Triton X-100, and the production of retinol was quantified by HPLC;Regulation of hepatic REH was studied by determining the relationship between its activity and physiological indicators of vitamin A nutritional status. Maximally stimulated hydrolase activity was not strongly influenced by serum or liver vitamin A concentrations in rats or pigs. However, in reduced detergent and substrate concentrations, REH activity was 3-fold higher (p < 0.01) in rats with depleted liver reserves (<10 (mu)g/g). Thus, REH may play a role in the homeostatically controlled release of vitamin A from liver, by mechanisms that are masked in vitro by concentrated detergent;Bile salts also strongly affected the level and interanimal variability of REH activity. Notably, rat REH activity in individual livers was 10-fold higher in CHAPS compared to sodium cholate, and less variable (5- compared to 37-fold). Therefore, the variability others have reported may be due to the use of sodium cholate to stimulate REH;13-cis, 9-cis and 9,13-cis Retinyl palmitate were excellent substrates for pig liver REH, being hydrolyzed at rates 2- to 7-times greater than the all-trans isomer with similar apparent K(,m) values and pH dependencies. However, by salt fractionation and column chromatography, three enzyme preparations were separated, which differed in their sensitivities to inhibitors and in their detergent-dependent substrate specificities. Thus, retinyl ester hydrolysis in the liver may well involve more than one enzyme.
DOI
https://doi.org/10.31274/rtd-180813-6881
Publisher
Digital Repository @ Iowa State University, http://lib.dr.iastate.edu/
Copyright Owner
Dale Alan Cooper
Copyright Date
1986
Language
en
Proquest ID
AAI8703697
File Format
application/pdf
File Size
134 pages
Recommended Citation
Cooper, Dale Alan, "Properties of retinyl ester hydrolase activity in pig and rat liver " (1986). Retrospective Theses and Dissertations. 8238.
https://lib.dr.iastate.edu/rtd/8238
