Akihiro Inoue

Cloning and sequence analysis of cDNA encoding the precursor of a human endothelium-derived vasoconstrictor peptide, endothelin: Identity of human and porcine endothelin

A cDNA encoding a human endothelium‐derived vasoconstrictor peptide, endothelin, was isolated from a human placenta cDNA library. The nucleotide sequence of this cDNA clone showed that the primary structure of the human preproendothelin has 212 amino acid residues and is highly homologous to porcine preproendothelin, and that human endothelin is identical with porcine endothelin.

cDNA cloning, sequence analysis and tissue distribution of rat preproendothelin-1 mRNA

We report the cloning of a full-length cDNA encoding rat preproendothelin-1 (preproET-1). The predicted rat preproET-1 consists of 202 amino acid residues and highly similar to human, porcine and bovine preproET-1, respectively. The deduced 21-residue sequence of mature rat ET-1 is identical to human, porcine, canine and bovine ET-1. As in other mammalian species, the mature ET-1 is predicted to be produced from a 39-residue big ET-1 in the rat. Northern blot analysis showed that a single 2.3-kb preproET-1 mRNA is expressed not only in vascular endothelial cells but also in other rat tissues, including the lung, brain, uterus, stomach, heart, adrenal gland and kidney. These findings suggest that ET-1 may play roles as a local mediator in multiple organs both within and outside the cardiovascular system in the rat.

The human preproendothelin-1 gene: Possible regulation by endothelial phosphoinositide turnover signaling

Summary Preproendothelin-1 (ppET-1) mRNA has previously been demonstrated to be markedly induced in cultured endothelial cells (EC) by the addition to the medium of thrombin, an agent known to stimulate phosphoinositide turnover in EC. In this study, the mechanism of regulation of ppET-1 mRNA expression was investigated in cultured human umbilical vein EC by RNA blot analysis with cloned ppET-1 gene as a probe. The mRNA for ppET-1 was rapidly upregulated by O-tetradecanoylphorbol-13-acetate (TPA) (0.5

A novel regulatory effect of myosin light chain kinase from smooth muscle on the ATP-dependent interaction between actin and myosin.

mUM) and by ionomycin (5

Differential tissue expression of multiple genes for chicken smooth muscle/nonmuscle myosin regulatory light chains

mUM) within 10 min of addition to the medium, but not by forskolin (50

The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes

mUM). The rapid induction of ppET-1 mRNA by TPA or ionomycin occurred even in the presence of cycloheximide, indicating that the mRNA induction does not require de novo protein synthesis. The ppET-1 mRNA was an extremely unstable species of mRNA with an apparent half-life of about 15 min. However, the half-life of ppET-1 mRNA was not appreciably affected by TPA or ionomycin, suggesting that the mRNA induction by these agents is mostly due to an activation of the transcription of the mRNA. These observations indicate that the production of ET-1 in human EC can be controlled by a transcriptional gene regulation directly coupled to the intracellular signals from the phosphoinositide-turnover pathway, i.e., activation of protein kinase C and increase in intracellular Ca2+. These mechanisms are discussed in relation to information on the primary structure of cloned ppET-1 gene

Primary structure, synthesis, and biological activity of rat endothelin, an endothelium-derived vasoconstrictor peptide

The actin-binding activity of myosin light chain kinase (MLCK) from smooth muscle was studied with special reference to the ATP-dependent interaction between actin and myosin. MLCK in the presence of calmodulin endowed sensitivity to Ca2+ on the movement of actin filaments on phosphorylated myosin from smooth muscle that was fixed on a coverslip. This regulatory effect was not attributable to the kinase activity of MLCK but could be explained by its actin-binding activity. The importance of the actin-binding activity was further substantiated by results of an experiment with Nitellopsis actin-cables in which MLCK regulated the interaction under conditions where MLCK was exclusively associated with the actin-cables.

The human preproendothelin-1 gene. Complete nucleotide sequence and regulation of expression.

The cDNA clones for two distinct mRNAs encoding one of the two known isoforms of chicken smooth muscle/nonmuscle myosin regulatory light chain were isolated. The nucleotide sequences of these cDNAs were very similar to each other (99% nucleotide identities) in the 516 bp translated regions and in the first 33 bp of the 3 noncoding regions, whereas the rest of the 3 noncoding regions and the 5 noncoding regions had no significant similarity. Genomic Southern blot analysis showed that these two mRNAs were encoded in two individual genes. Whereas these two genes encoded almost identical polypeptides with only one conservative substitution of amino acid residues, expression of the mRNAs was differentially regulated both at the transcriptional and translational levels in various tissues of the chicken.