Age-related changes in catalytic activity, enzyme mass, mRNA, and subcellular distribution of hepatic neutral cholesterol ester hydrolase in female rats.
Activity and protein mass of hepatic neutral cholesteryl ester hydrolase (CEH) were measured in liver cytosol and washed microsomes of female Sprague-Dawley rats aged 3, 4, 7, 9, 13, and 16 wk. CEH mRNA was also measured. The microsomal component varied with age and contributed a greater fraction of total activity in females than previously reported in males. Nevertheless, the cytosolic component accounted for 62-80% of activity and 77-94% of immunoreactive protein in postmitochondrial fractions. Cytosolic and microsomal CEH specific activities, relative to total protein, decreased 94 and 83%, respectively, from 3 to 4 wk, prior to onset of puberty at 5 wk, and increased 360 and 137%, respectively, from 12 to 16 wk. These results contrast with an earlier study, in which cytosolic CEH activity of males increased with puberty and declined after 12 wk. Although cytosolic CEH was activated by protein kinase A and inhibited by alkaline phosphatase treatment at all ages, protein kinase activation peaked at 4 wk, coinciding with the initial decrease in specific activity. Specific activity in cytosol and microsomes correlated with CEH mass at all ages, suggesting that this CEH accounts for most variation in cellular activity. In contrast, CEH mRNA varied little from 3-16 wk, indicating that transcriptional regulation does not make a major contribution to the variation in CEH activity and mass in females, although it may make an important contribution to male-female differences in CEH expression. Specific activities of cytosolic and microsomal CEH, relative to immunoreactive CEH protein mass, exhibited changes consistent with posttranslational regulation. These results indicate gender-specific multivalent regulation of hepatic CEH by posttranslational mechanisms during development of female rats.
Natarajan
R
R
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0614, USA.
Ghosh
S
S
Grogan
W M
WM
D000375:Aging / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D006706:Homeostasis; D008099:Liver / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012044:Regression Analysis; D012737:Sex Factors; D012741:Sexual Maturation; D002787:Sterol Esterase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D017868:Cyclic AMP-Dependent Protein Kinases; D002787:Sterol Esterase; D000469:Alkaline Phosphatase