Inhibition of the tumor necrosis factor-α-converting enzyme by its pro domain

Patricia E. Gonzales, Ariel Solomon, Ann B. Miller, M. Anthony Leesnitzer, Irit Sagis, Marcos E. Milla

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

98 Citas (Scopus)

Resumen

Tumor necrosis factor-α-converting enzyme (TACE) is a disintegrin metalloproteinase that processes tumor necrosis factor and a host of other ectodomains. TACE is biosynthesized as a zymogen, and activation requires the removal of an inhibitory pro domain. Little is known about how the pro domain exerts inhibition for this class of enzymes. To study the inhibitory properties of the pro domain of TACE, we have expressed it in isolation from the rest of the protease. Here we show that the TACE pro domain (TACE Pro) is a stably folded protein that is able to inhibit this enzyme. TACE Pro inhibited the catalytic domain of TACE with an IC50. of 70 nM. In contrast, this inhibitory potency decreased over 30-fold against a TACE form containing the catalytic plus disintegrin/cysteine-rich domains (IC50 greater that 2 μM). The disintegrin/cysteine-rich region in isolation also decreases the interaction of TACE Pro with the catalytic domain. Surprisingly, we found that the cysteine switch motif located in TACE Pro was not essential for inhibition of the enzymatic activity of TACE; the pro domain variant C184A showed the same inhibitory potency against both TACE forms as wild type TACE Pro. X-ray absorption spectroscopy experiments indicate that binding of TACE Pro to the catalytic domain does include ligation of the catalytic zinc ion via the sulfur atom of its conserved Cys184 residue. Moreover, the binding of TACE Pro to the catalytic zinc ion partially oxidizes the catalytic zinc ion of the enzyme. Despite this, the nature of the interaction between the pro and catalytic domains of TACE is not consistent with a simple competitive model of inhibition based on cysteine switch ligation of the zinc ion within the active site of TACE.

Idioma originalInglés
Páginas (desde-hasta)31638-31645
Número de páginas8
PublicaciónJournal of Biological Chemistry
Volumen279
N.º30
DOI
EstadoPublicada - 23 jul. 2004
Publicado de forma externa

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