© 2000 American Heart Association, Inc.
Cellular Biology |
ADP-Ribosyl Cyclase in Rat Vascular Smooth Muscle Cells
Properties and Regulation
From the Department of Physiology and Biophysics, Division of Nephrology and Internal Medicine (F.G.S.d.T., J.C., M.L., T.P.D.), and Departments of Anesthesiology and Department of Medicine (E.N.C.), Mayo Clinic and Foundation, Mayo Medical School, Rochester, Minn.
Correspondence to Eduardo N. Chini, MD, PhD, Mayo Clinic, 200 First St, SW, Guggenheim 9, Rochester, MN 55905. E-mail chini.eduardo{at}mayo.edu
Abstract—We investigated whether
ADP-ribosyl cyclase (ADPR-cyclase) in rat vascular smooth muscle cells
(VSMCs) has enzymatic properties that differ from the
well-characterized CD38-antigen ADPR-cyclase, expressed in HL-60 cells.
ADPR-cyclase from VSMCs, but not CD38 ADPR-cyclase from HL-60 cells,
was inhibited by gangliosides (10 µmol/L) GT1B,
GD1, and GM3. Preincubation of membranes from
CD38 HL-60 cells, but not from VSMCs, with anti-CD38 antibodies
increased ADPR-cyclase activity; CD38 antigen was detected both in
VSMCs and in HL-60 cells. ADPR-cyclase in VSMC membranes was more
sensitive than CD38 HL-60 ADPR-cyclase to inactivation by
N-endoglycosidase F and to thermal inactivation at
45°C. The specific activity of ADPR-cyclase in membranes from VSMCs
was >20-fold higher than in membranes from CD38 HL-60 cells. Most
importantly, VSMC ADPR-cyclase was inhibited by Zn2+ and
Cu2+ ions; the inhibition by Zn2+ was dose
dependent, noncompetitive, and reversible by EDTA. In contrast,
Zn2+ stimulated the activity of CD38 HL-60 ADPR-cyclase and
other known types of ADPR-cyclases. Retinoids act either via the
nuclear receptor retinoic acid receptor or retinoid X receptor,
including all-trans retinoic acid (atRA), and panagonist
9-cis-retinoic acid–upregulated VSMC ADPR-cyclase; the
stimulatory effect of atRA was blocked by actinomycin D and
cycloheximide. 1,25(OH)2–Vitamin D3
(calciferol) stimulated VSMC ADPR-cyclase dose dependently at
subnanomolar concentrations (ED50
56 pmol/L).
Oral administration of atRA to rats resulted in an increase of
ADPR-cyclase activity in aorta (+60%) and, to a lesser degree, in
myocardium of left ventricle (+18%), but atRA had no
effect on ADPR-cyclases in lungs, spleen, intestinal smooth muscle,
skeletal muscle, liver, or testis. Administration of
3,5,3'-triiodothyronine (T3) to rats resulted in an
increase of ADPR-cyclase activity in aorta (+89%), but not in liver
or brain. We conclude the following: (1) ADPR-cyclase in VSMCs has
enzymatic properties distinct from "classic" CD38 ADPR-cyclase,
especially sensitivity to inhibition by Zn2+ and
Cu2+; (2) ADPR-cyclase in VSMCs is upregulated by various
retinoids, calcitriol, and T3 in vitro; and (3)
administration of atRA and T3 increases ADPR-cyclase in
aorta in vivo. We suggest that the cADPR signaling system plays an
important role in the regulation of VSMC functions in response to
steroid superfamily hormones.
Key Words: vascular smooth muscle cells • calciferols • antibodies • ADP-ribose • retinoids
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