TY - JOUR
T1 - Incorporation of steroidogenic pathways which produce cortisol and aldosterone from cholesterol into nonsteroidogenic cells
AU - Mathew, Porunelloor A.
AU - Mason, J. Ian
AU - Trant, John M.
AU - Waterman, Michael R.
N1 - Funding Information:
Theses tudiesw eres upportedin part by USPHS research grants GM37942, AGO8175, and GM16488.P .A.M. was supportedb y Grant I-624 from The Robert A. Welch Foundationa nd J.M.T. was supported by USPHS Training Grant 5-T32HDO7190W. e greatlya ppreciatet he editorial assistanceof SusanA lexander. Wael so appreciate the generosityo f MatthewL orence of this institution in providing us with the expressionv ector containing cDNA insert specific for human placental3 #LHSD.
PY - 1990/10/1
Y1 - 1990/10/1
N2 - Cortisol production from cholesterol requires the activity of four steroid hydroxylases: cholesterol side chain cleavage cytochrome P-450 (P-450scc), 17α-hydroxylase cytochrome P-450 (P-45017α), 21-hydroxylase cytochrome P-450 (P-450c21) and 11β-hydroxylase cytochrome P-450 (P-45011β). We have previously shown that transformed, nonsteroidogenic COS 1 cells derived from monkey kidney are a useful system for expression of various forms of cytochrome P-450. The present study shows that COS 1 cell cultures multiply transfected with six plasmids containing all four steroid hydroxylases, 3β-hydroxysteroid dehydrogenase /Δ5→4-isomerase (3βHSD) and adrenodoxin produce cortisol and aldosterone when 22(R)-hydroxycholesterol is supplied to the system. When pregnenolone is used as substrate, various intermediate metabolites are detected at different time points further establishing the incorporation of complete functional steroidogenic pathways into the nonsteroidogenic cell cultures. Since the first and the last reactions in these pathways take place in the mitochondrion, the movement of various intermediate metabolites from mitochondrion to endoplasmic reticulum and back to mitchondrion occurs in and between COS 1 cells.
AB - Cortisol production from cholesterol requires the activity of four steroid hydroxylases: cholesterol side chain cleavage cytochrome P-450 (P-450scc), 17α-hydroxylase cytochrome P-450 (P-45017α), 21-hydroxylase cytochrome P-450 (P-450c21) and 11β-hydroxylase cytochrome P-450 (P-45011β). We have previously shown that transformed, nonsteroidogenic COS 1 cells derived from monkey kidney are a useful system for expression of various forms of cytochrome P-450. The present study shows that COS 1 cell cultures multiply transfected with six plasmids containing all four steroid hydroxylases, 3β-hydroxysteroid dehydrogenase /Δ5→4-isomerase (3βHSD) and adrenodoxin produce cortisol and aldosterone when 22(R)-hydroxycholesterol is supplied to the system. When pregnenolone is used as substrate, various intermediate metabolites are detected at different time points further establishing the incorporation of complete functional steroidogenic pathways into the nonsteroidogenic cell cultures. Since the first and the last reactions in these pathways take place in the mitochondrion, the movement of various intermediate metabolites from mitochondrion to endoplasmic reticulum and back to mitchondrion occurs in and between COS 1 cells.
KW - COS 1 cells
KW - Multiple transfection
KW - Steroid hydroxylases
UR - http://www.scopus.com/inward/record.url?scp=0025075658&partnerID=8YFLogxK
U2 - 10.1016/0303-7207(90)90046-B
DO - 10.1016/0303-7207(90)90046-B
M3 - Article
C2 - 2292341
AN - SCOPUS:0025075658
SN - 0303-7207
VL - 73
SP - 73
EP - 80
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
IS - 1
ER -