Role of chloride ions in modulation of the interaction between von Willebrand factor and ADAMTS-13

Raimondo De Cristofaro, Flora Peyvandi, Roberta Palla, Silvia Lavoretano, Rossana Lombardi, Giuliana Merati, Federica Romitelli, Enrico Di Stasio, Pier Mannuccio Mannucci

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The degradation of von Willebrand factor (VWF) depends on the activity of a zinc protease (referred to as ADAMTS-13), which cleaves VWF at the Tyr 1605-Met1606 peptide bond. Little information is available on the physiological mechanisms involved in regulation of AD-AMTS-13 activity. In this study, the role of ions on the ADAMTS-13/VWF interaction was investigated. In the presence of 1.5 M urea, the protease cleaved multimeric VWF in the absence of NaCl at pH 8.00 and 37 °C, with an apparent k cat/Km ≅ 3.4 × 104 M-1 S-1, but this value decreased by ∼10-fold in the presence of 0.15 M NaCl. Using several monovalent salts, the inhibitory effect was attributed mostly to anions, whose potency was inversely related to the corresponding Jones-Dole viscosity B coefficients (ClO4 - > Cl - > F-). The specific inhibitory effect of anions was due to their binding to VWF, which caused a conformational change responsible for quenching the intrinsic fluorescence of the protein and reducing tyrosine exposition to bulk solvent. Ristocetin binding to VWF could reduce the apparent affinity and reverse the inhibitory effect of chloride. We hypothesize that, after secretion into the extracellular compartment, VWF is bound by chloride ions abundantly present in this milieu, becoming unavailable to proteolysis by AD-AMTS-13. Shear forces, which facilitate GpIbα binding (this effect being artificially obtained by ristocetin), can reverse the inhibitory effect of chloride, whose concentration gradient across the cell membrane may represent a simple but efficient strategy to regulate the enzymatic activity of ADAMTS-13.

Original languageEnglish
Pages (from-to)23295-23302
Number of pages8
JournalJournal of Biological Chemistry
Issue number24
Publication statusPublished - Jun 17 2005

ASJC Scopus subject areas

  • Biochemistry


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