1. Molecular identification and characterization of novel membrane-bound metalloprotease, the soluble secreted form of which hydrolyzes a variety of vasoactive peptides
One class of zinc metalloproteases, represented by neutral endopeptidase 24.11 and endothelin-converting enzyme, has been shown to be involved in proteolytic activation or inactivation of many regulatory peptides. Here, we report molecular cloning and characterization of a novel member of this type II membrane-bound metalloprotease family, termed soluble secreted endopeptidase (SEP). Alternative splicing results in the generation of another transcript, SEPΔ, which lacks a 69-base pair nucleotide segment following the transmembrane helix. Both SEP and SEPΔmRNA are detected in all mouse tissues examined. Transfection of an SEP cDNA expression construct resulted in the expression of the membrane-bound form of SEP in the early secretory pathway as well as the soluble secreted form of the enzyme in the culture medium. In contrast, transfection of the SEPΔ cDNA only results in the expression of the membrane-bound form. In vitroenzymological analysis of the recombinant soluble form of SEP demonstrated that it hydrolyzes a variety of vasoactive peptides, including endothelin-1, atrial natriuretic peptide, and angiotensin I. This activity of SEP was inhibited by phosphoramidon and the neutral endopeptidase 24.11 specific inhibitor thiorphan, but it was only partially inhibited by the endothelin-converting enzyme specific inhibitor FR901533. These findings suggest that SEP is a novel metalloprotease that possesses a broad substrate specificity and that it may be involved in the metabolism of biologically active peptides intracellulary as well as extracellularly.
2. Constitutive lysosomal targeting and degradation of bovine endothelin-converting enzyme-1a mediated by novel signals in its alternatively spliced cytoplasmic tail
Abstrak : Endothelin-converting enzyme-1 (ECE-1) is a type II membrane protein that catalyzes the proteolytic activation of big endothelin-1 to endothelin-1 (ET-1). The subcellular distribution of ECE-1, and hence the exact site of physiological activation of big ET-1, remains controversial. Here, we demonstrate with several complementary methods that the two alternatively spliced bovine ECE-1 isoforms, ECE-1a and ECE-1b, differing only in the first 30 amino acids of their N-terminal cytoplasmic tails, exhibit strikingly distinct intracellular sorting patterns. Bovine ECE-1a, which is responsible for the intracellular cleavage of big ET-1 in endothelial cells, is constitutively recruited into the lysosome, where it is rapidly degraded. In contrast, bovine ECE-1b, the isoform found in cultured smooth muscle cells, is transported to the plasma membrane by a default pathway and functions as an ectoenzyme. Mutational analyses reveal that the N-terminal tip of the cytoplasmic domain of bovine ECE-1a contains novel proline-containing signals that mediate constitutive lysosomal targeting. Analyses of chimeric ECE-1/transferrin receptors demonstrate that the cytoplasmic tail of bovine ECE-1a is sufficient for the lysosomal delivery and rapid degradation. Our results suggest that the distinct intracellular targeting of bovine ECE-1 isoforms may provide new insights into functional aspect of the endothelin system and that the cell permeability of ECE inhibitor compounds should be carefully considered during their pharmacological development.
3. Consecutive administration of paraquat to rats induces enhanced cholesterol peroxidation and lung injury
4. Accidental hanging by a sweater: an unusual case
5. Nasopharyngeal carcinoma in Indonesia has a low prevalence of the 30-base pair deletion of Epstein-Barr virus latent membrane protein
6. 7-Hydroperoxycholesterol and oxysterols as indices of oxidative stress: chronic ethanol feeding and rat skeletal muscle
7. Perbandingan korelasi penentuan tinggi badan antara metode pengukuran panjang tibia perkutaneus dan panjang telapak kaki
8. Membrane-bound metalloprotease and soluble secreted form thereof
9. Perbandingan korelasi antara metode pengukuran panjang bahu dan panjang lengan atas perkutaneus terhadap panjang badan jenazah
10. Molecular cloning and characterization of a novel metalloprotease that cleaves big endothelin-1 to produce active endothelin-1
11. PERTUMBUHAN LARVA LALAT PADA WISTAR RATTUS NOVERGICUS AKIBAT PENGGUNAAN DOSIS LETAL KLORPROMAZIN SEBAGAI PERKIRAAN POST MORTEM INTERVAL
12. Perbandingan Analisis Paternity Index Pada Kasus Ragu Ayah (Disputed Paternity)
13. KEMATIAN AKIBAT LUKA TEMBAK YANG DIOTOPSI DI INSTALASI KEDOKTERAN FORENSIK RSUP Dr. SARDJITO YOGYAKARTA TAHUN 1993-2013
14. VALIDITAS HASIL PEMERIKSAAN METODE AGLUTINASI DIREK DAN ELUSI ABSORPSI UNTUK IDENTIFIKASI GOLONGAN DARAH PADA JENAZAH
15. Hubungan Sebab Kematian dengan Alkohol pada Jenazah Forensik di Instalasi Kedokteran Forensik RSUP Dr. Sardjito Tahun 1993-2013
16. UJI DIAGNOSTIK PENENTUAN JENIS KELAMIN METODE PEMERIKSAAN DRUMSTICK NEUTROFIL DIBANDINGKAN DENGAN METODE PEMERIKSAAN AMELOGENIN DNA (DEOXYRIBONUCLEIC ACID)
17. PERBANDINGAN TANDA-TANDA PATOLOGIS YANG DITEMUKAN PADA JENAZAH KORBAN KECELAKAAN LALU LINTAS ANTARA KORBAN PEMAKAI HELM DAN BUKAN PEMAKAI HELM
18. PENDETEKSIAN GOLONGAN DARAH ABO DENGAN SAMPEL TULANG JENAZAH KORBAN KONDISI TERBAKAR MENGGUNAKAN METODE LACAK BIOLOGI MOLEKULER
19. ANALISA GENETIK LOKUS TH01, TPO, DAN CFS1PO SHORT TANDEM REPEATS PADA PENDUDUK YOGYAKARTa
20. Kajian surat keterangan kematian yang dikeluarkan Instalasi Kedokteran Forensik Rumah Sakit Dr. Sardjito Yogyakarta
21. Three sudden death cases with a suicide note
22. Isolation and enzymological characterization of a novel soluble, secreted metalloprotease that hydrolyzes atrial natriuretic peptide and bradykinin