Zinc induces exposure of hydrophobic sites in the C-terminal domain of gC1q-R/p33.
Endothelial cells and platelets are known to express gC1q-R on their surface. In addition to C1q, endothelial cell gC1q-R has been shown to bind high molecular weight kininogen (HK) and factor XII (FXII). However, unlike C1q, whose interaction with gC1q-R does not require divalent ions, the binding of HK to gC1q-R is absolutely dependent on the presence of zinc. However, the mechanism by which zinc modulates this interaction is not fully understood. To investigate the role of zinc, binding studies were done using the hydrophobic dye, bis-ANS. The fluorescence intensity of bis-ANS, greatly increases and the emission maximum is blue-shifted from 525 to 485nm upon binding to hydrophobic sites on proteins. In this report, we show that a blue-shift in emission maximum is also observed when bis-ANS binds to gC1q-R in the presence but not in the absence of zinc suggesting that zinc induces exposure of hydrophobic sites in the molecule. The binding of bis-ANS to gC1q-R is specific, dose-dependent, and reversible. In the presence of zinc, this binding is abrogated by monoclonal antibody 74.5.2 directed against gC1q-R residues 204-218. This segment of gC1q-R, which corresponds to the beta6 strand in the crystal structure, has been shown previously to be the binding site for HK. A similar trend in zinc-induced gC1q-R binding was also observed using the hydrophobic matrix octyl-Sepharose. Taken together, our data suggest that zinc can induce the exposure of hydrophobic sites in the C-terminal domain of gC1q-R involved in binding to HK/FXII.
Kumar
Rajeev
R
Department of Medicine, State University of New York, Stony Brook, NY 11794-8161, USA.
Peerschke
Ellinor I B
EI
Ghebrehiwet
Berhane
B
D001665:Binding Sites; D002352:Carrier Proteins; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors; D057927:Hydrophobic and Hydrophilic Interactions; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D011487:Protein Conformation; D011951:Receptors, Complement / Q000737:chemistry* / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology*
C087299:C1QBP protein, human; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D019679:Kininogen, High-Molecular-Weight; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D011951:Receptors, Complement; C032402:complement 1q receptor; D015032:Zinc