Intracellular Activation of Human Adamalysin 19/Disintegrin and Metalloproteinase 19 by Furin Occurs via One of the Two Consecutive Recognition Sites

• Published in: The Journal of biological chemistry Vol. 277; no. 28; pp. 25583 - 25591
• Main Authors: Kang, Tiebang, Zhao, Yun-Ge, Pei, Duanqing, Sucic, Joseph F., Sang, Qing-Xiang Amy
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 12.07.2002; American Society for Biochemistry and Molecular Biology
• Subjects: ADAM Proteins; Amino Acid Sequence; Animals; Base Sequence; Cell Line; Cell Membrane - enzymology; Cytoplasm - enzymology; Disintegrins; DNA Primers; Endoplasmic Reticulum - enzymology; Endoplasmic Reticulum - metabolism; Enzyme Activation; Furin; Golgi Apparatus - enzymology; Golgi Apparatus - metabolism; Humans; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Metalloendopeptidases; Metalloproteases; Muscle Proteins - chemistry; Muscle Proteins - genetics; Muscle Proteins - metabolism; Mutagenesis; Protein Processing, Post-Translational; Subtilisins - physiology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M203532200

Adamalysin 19 (a disintegrin and metalloproteinase 19, ADAM19, or meltrin β) is a plasma membrane metalloproteinase. Human ADAM19 zymogen contains two potential furin recognition sites (RX(K/R)R),196KRPR200Rand199RRMK203R, between its pro- and catalytic domains. Protein N-terminal sequencing revealed that the cellular mature forms of hADAM19 started at204EDLNSMK, demonstrating that the preferred furin cleavage site was the200RMK203R↓204EDLN. Those mature forms were catalytically active. Both Pittsburgh mutant of α1-proteinase inhibitor and dec-Arg-Val-Lys-Arg-chloromethyl ketone, two specific furin inhibitors, blocked the activation of hADAM19. Activation of hADAM19 was also blocked by brefeldin A, which inhibits protein trafficking from the endoplasmic reticulum to the Golgi, or A23187, a calcium ionophore known to inhibit the autoactivation of furin. When 202KR were mutated to AA, the proenzyme was also activated, suggesting that197RPRR is an alternative activation site. Furthermore, only pro-forms of hADAM19 were detected in the 199RR to AA mutant, which abolished both furin recognition sites. Moreover, the zymogens were not converted into their active forms in two furin-deficient mammalian cell lines; co-expression of hADAM19 and furin in these two cell lines restored zymogen activation. Finally, co-localization between furin and hADAM19 was identified in the endoplasmic reticulum-Golgi complex and/or the trans-Golgi network. This report is the first thorough investigation of the intracellular activation of adamalysin 19, demonstrating that furin activated pro-hADAM19 in the secretory pathway via one of the two consecutive furin recognition sites.