Subcellular localization of cyclic ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities in porcine airway smooth muscle

Thomas A. White, Sonja Johnson, Timothy F. Walseth, Hon Cheung Lee, Richard M. Graeff, Cyrus B. Munshi, Y.s. Prakash, Gary C Sieck, Mathur S. Kannan

Research output: Contribution to journalArticle

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Abstract

Recent studies have provided evidence for a role of cyclic ADP-ribose (cADPR) in the regulation of intracellular calcium in smooth muscles of the intestine, blood vessels and airways. We investigated the presence and subcellular localization of ADP-ribosyl cyclase, the enzyme that catalyzes the conversion of β-NAD+ to cADPR, and cADPR hydrolase, the enzyme that degrades cADPR to ADPR, in tracheal smooth muscle (TSM). Sucrose density fractionation of TSM crude membranes provided evidence that ADP-ribosyl cyclase and cADPR hydrolase activities were associated with a fraction enriched in 5'-nucleotidase activity, a plasma membrane marker enzyme, but not in a fraction enriched in either sarcoplasmic endoplasmic reticulum calcium ATPase or ryanodine receptor channels, both sarcoplasmic reticulum markers. The ADP-ribosyl cyclase and cADPR hydrolase activities comigrated at a molecular weight of approximately 40 kDa on SDS-PAGE. This comigration was confirmed by gel filtration chromatography. Investigation of kinetics yielded K(m) values of 30.4 ± 1.5 and 695.3 ± 171.2 μM and V(max) values of 330.4 ± 90 and 102.8 ± 17.1 nmol/mg/h for ADP-ribosyl cyclase and cADPR hydrolase, respectively. These results suggest a possible role for cADPR as an endogenous modulator of [Ca2+](i) in porcine TSM cells. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)64-71
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1498
Issue number1
DOIs
StatePublished - Oct 20 2000

Fingerprint

ADP-ribosyl Cyclase
Smooth Muscle
Swine
Cyclic ADP-Ribose
Enzymes
Sarcoplasmic Reticulum Calcium-Transporting ATPases
Calcium-Sensing Receptors
5'-Nucleotidase
Ryanodine Receptor Calcium Release Channel
Sarcoplasmic Reticulum
Endoplasmic Reticulum
NAD
Smooth Muscle Myocytes
Intestines
Gel Chromatography
Blood Vessels
Sucrose
Polyacrylamide Gel Electrophoresis
Molecular Weight
Cell Membrane

Keywords

  • ADP-ribosyl cyclase
  • Airway smooth muscle
  • Cyclic ADP-ribose
  • Cyclic ADP-ribose hydrolase

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Biophysics

Cite this

Subcellular localization of cyclic ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities in porcine airway smooth muscle. / White, Thomas A.; Johnson, Sonja; Walseth, Timothy F.; Lee, Hon Cheung; Graeff, Richard M.; Munshi, Cyrus B.; Prakash, Y.s.; Sieck, Gary C; Kannan, Mathur S.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1498, No. 1, 20.10.2000, p. 64-71.

Research output: Contribution to journalArticle

White, Thomas A. ; Johnson, Sonja ; Walseth, Timothy F. ; Lee, Hon Cheung ; Graeff, Richard M. ; Munshi, Cyrus B. ; Prakash, Y.s. ; Sieck, Gary C ; Kannan, Mathur S. / Subcellular localization of cyclic ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities in porcine airway smooth muscle. In: Biochimica et Biophysica Acta - Molecular Cell Research. 2000 ; Vol. 1498, No. 1. pp. 64-71.
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AU - Graeff, Richard M.

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