{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T15:17:34Z","timestamp":1759936654820,"version":"build-2065373602"},"reference-count":29,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2010,7,22]],"date-time":"2010-07-22T00:00:00Z","timestamp":1279756800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Anatomical Record"],"published-print":{"date-parts":[[2010,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The process of regulation of NOS after production of nitric oxide is not yet delineated. Protein kinase G may exert a feedback regulation of this enzyme. We used diaminofluorescein assays to detect changes in basal nitric oxide production caused by modulators of protein kinase G activity in freshly isolated ovine lung microvascular endothelial cells. We also used fluorescence activated cell sorter analysis (FACS) to determine molecular and phosphorylation changes caused by PKG activation with 8\u2010Br\u2010cGMP. The PKG activator, 8\u2010Br\u2010cGMP (100 \u03bcM) produced a shift in the basal NO production curve downward. The inhibition began within 5 min and was sustained over 4.5 hr. The two protein kinase G inhibitors 100 \u03bcM Rp\u20108\u2010Br\u2010PET\u2010cGMPS and 50 nM guanosine 3\u2032\u20105\u2032\u2010cyclic monophosphoro thionate\u20108\u2010Br\u2010Rp isomer Na salt and the cGMP inhibitor 4 \u03bcM Rp\u20108\u2010pCPT\u2010cGMPS all enhanced NO production as seen by the upward shift in the basal NO curve. Conversely, the PKG activator drug, 100 \u03bcM guanosine\u20103\u2032\u20105\u2032\u2010cyclic monophosphate\u2010\u03b2\u2010phenyl\u20101NF\u2010ethano\u20108\u2010bromo sodium salt decreased NO production causing a downward shift in the basal curve. FACS analysis revealed that 5 \u03bcM 8\u2010Br\u2010cGMP in &lt;5 min caused an increase in N\u2010terminal labeling of NOS and a decrease in both C\u2010terminal and serine 1177 labeling of NOS. 8\u2010Br\u2010cGMP appeared to increase PKG 1\u03b1 and to decrease PKG 1\u03b2 labeling. Changes in other phosphorylation sites were less consistent but overall mean channel fluorescence increased from 19.92 to 217.36 for serine 116 and decreased from 329.27 to 254.03 for threonine 495 phosphorylation. Data indicated that PKG caused both molecular and phosphorylation changes in NOS. Anat Rec 293:1755\u20131765, 2010. \u00a9 2010 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/ar.21169","type":"journal-article","created":{"date-parts":[[2010,7,22]],"date-time":"2010-07-22T12:00:55Z","timestamp":1279800055000},"page":"1755-1765","source":"Crossref","is-referenced-by-count":1,"title":["A Fluorescence\u2010Activated Cell Sorter Analysis of the Relationship Between Protein Kinase G and Endothelial Nitric Oxide Synthase"],"prefix":"10.1002","volume":"293","author":[{"given":"Theresa A.","family":"John","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. 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