The source of heme for vascular heme oxygenase II: de novo heme biosynthesis in rat aorta.
Heme is an essential prosthetic group or substrate for many proteins, including hemoglobin, and hemo enzymes such as nitric oxide synthase, soluble guanylyl cyclase, and heme oxygenase (HO). HO is responsible for the breakdown of heme into equimolar amounts of biliverdin, iron, and carbon monoxide, the latter of which is thought to play a role in the regulation of vascular tone. It is not clear whether the source of heme for cardiovascular functions is derived from uptake from the extracellular milieu or synthesis. In this study, we tested the hypothesis that blood vessels obtain their supply of heme for HO through de novo synthesis. Adult male Sprague-Dawley rat aorta was incubated at 37 degrees C in Krebs' solution with 1 micro M [14C]delta-aminolevulinic acid (ALA). [14C]ALA uptake was linear for about 30 min and reached a plateau at approximately 100 min. The radioactivity was incorporated into porphyrins and heme as determined by esterification of 14C-labelled metabolites and thin-layer chromatography. The first and rate-limiting step of heme biosynthesis is catalyzed by ALA synthase (ALA-S), the activity of which was determined in rat aorta using a radiometric assay, approximately 250 nmol x (g wet mass)(-1) x h(-1). Inducing HO-1 in rat aorta with S-nitroso-N-acetylpenicillamine (500 micro M) did not increase ALA-S activity as compared with basal activity levels of the enzyme. It appears that there is a sufficient amount of heme available under basal ALA-S activity conditions to meet the increased demand for heme resulting from HO-1 induction. These observations indicate that the complete enzymatic pathway for de novo heme biosynthesis resides in rat aorta and furthermore indicate that de novo heme synthesis is capable of supplying a substantial portion of the heme substrate for HO in the aorta.
Jaronczyk
Kinga
K
Department of Pharmacology and Toxicology, Faculty of Health Sciences, Queen's University, Kingston, ON, Canada.
Bui
Loc
L
Soong
Jonathan M
JM
McLaughlin
Brian E
BE
Marks
Gerald S
GS
Brien
James F
JF
Nakatsu
Kanji
K
D000624:5-Aminolevulinate Synthetase / Q000378:metabolism / Q000494:pharmacology; D000622:Aminolevulinic Acid / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001011:Aorta / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002170:Canada; D002250:Carbon Radioisotopes; D005998:Glycine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006418:Heme / Q000096:biosynthesis* / Q000737:chemistry; D006419:Heme Oxygenase (Decyclizing) / Q000096:biosynthesis*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology
D002250:Carbon Radioisotopes; D006418:Heme; D026423:S-Nitroso-N-Acetylpenicillamine; D000622:Aminolevulinic Acid; D006419:Heme Oxygenase (Decyclizing); C105245:heme oxygenase-2; D000624:5-Aminolevulinate Synthetase; D005998:Glycine