Assembly of Weibel-Palade body-like tubules from N-terminal domains of von Willebrand factor.
Endothelial cells assemble von Willebrand factor (VWF) multimers into ordered tubules within storage organelles called Weibel-Palade bodies, and tubular packing is necessary for the secretion of VWF filaments that can bind connective tissue and recruit platelets to sites of vascular injury. We now have recreated VWF tubule assembly in vitro, starting with only pure VWF propeptide (domains D1D2) and disulfide-linked dimers of adjacent N-terminal D'D3 domains. Assembly requires low pH and calcium ions and is reversed at neutral pH. Quick-freeze deep-etch electron microscopy and three-dimensional reconstruction of negatively stained images show that tubules contain a repeating unit of one D'D3 dimer and two propeptides arranged in a right-handed helix with 4.2 units per turn. The symmetry and location of interdomain contacts suggest that decreasing pH along the secretory pathway coordinates the disulfide-linked assembly of VWF multimers with their tubular packaging.
Huang
Ren-Huai
RH
Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110, USA.
Wang
Ying
Y
Roth
Robyn
R
Yu
Xiong
X
Purvis
Angie R
AR
Heuser
John E
JE
Egelman
Edward H
EH
Sadler
J Evan
JE
D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D007477:Ions; D007834:Lasers; D008027:Light; D008854:Microscopy, Electron; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012542:Scattering, Radiation; D020302:Weibel-Palade Bodies / Q000737:chemistry* / Q000502:physiology; D014841:von Willebrand Factor / Q000737:chemistry*
D004220:Disulfides; D007477:Ions; D010455:Peptides; D014841:von Willebrand Factor