Expression in human endothelial cells of ADP-ribosylation factors, 20-kDa guanine nucleotide-binding proteins involved in the initiation of vesicular transport
Journal
Journal of Molecular and Cellular Cardiology
Journal Volume
28
Journal Issue
9
Pages
1911-1920
Date Issued
1996
Author(s)
Abstract
ADP-ribosylation factors (ARFs) are ~ 20-kDa, guanine nucleotide-binding proteins, initially discovered as stimulators of cholera toxin ADP-ribosyltransferase activity and subsequently shown to participate in vesicular trafficking. Five of the six mammalian ARFs have been identified in human tissues by molecular cloning. They fall into three classes (class I: ARFs 1-3; class II: ARFs 4, 5; class III: ARF 6) based on deduced amino acid sequence, size, phylogenetic analysis, and gene structure. Similar to the rab family of ~ 20 kDa guanine nucleotide-binding proteins, the ARFs appear to function in specific trafficking pathways. The presence of a specific ARF might serve as a marker for that pathway. To verify expression of ARF mRNA and protein in human umbilical vein endothelial cells, immunoreactivity using antibodies specific for each ARF class, quantitative polymerase chain reaction (PCR) using ARE-specific, internal cRNA standards containing unique restriction enzyme cleavage sites introduced by point mutations, and Northern analysis with probes specific for ARFs 1, and 3-6, were utilized. PCR and Northern analysis were in agreement in showing that amounts of mRNA for ARF 1 and ARF 4 were similar and higher than those of ARF 3 and ARF 5 which were greater than ARF 6. Primarily, Class I ARF proteins were detected by immunoreactivity, with the majority in the supernatant fraction. The relative expression of ARFs in endothelial cells thus differs from that in neuronal tissues where it had been found that ARF 3 is the predominant species.
Subjects
ADP-ribosylation factors; Guanine nucleotide-binding proteins; Human umbilical vein endothelial cells; Quantitative polymerase chain reaction
SDGs
Other Subjects
adenosine diphosphate ribosylation factor; complementary RNA; guanine nucleotide binding protein; messenger RNA; article; cell vacuole; controlled study; human; human cell; immunohistochemistry; Northern blotting; point mutation; polymerase chain reaction; priority journal; protein expression; restriction mapping; vascular endothelium; Mammalia
Publisher
Academic Press
Type
journal article