High molecular weight kininogen and factor XII binding to endothelial cells and astrocytes.
We have quantitated the binding of high molecular weight kininogen (HK) to human microvascular endothelial cells of lung and dermal origin as well as to astrocytes and compared the results with those reported for human umbilical vein endothelial cells (HUVEC). We also reassessed parameters of binding to HUVEC employing cells in suspension as well as cells attached to the culture plate and report similar numbers of sites varying from 6.96x10(5) to 7.71x10(5) per cell. The present study shows that HK binds with high specificity and affinity to microvascular endothelial cells (Kd = 1.86 to 4.5 nM) compared to HUVEC (Kd = 10.35 nM) but with lower affinity to astrocytes (Kd = 23.73 nM). Human cytokeratin 1, urokinase plasminogen activator receptor and gC1qR were found to be HK binding proteins present at the surface of microvascular endothelial cells and astrocytes analogous to that seen in HUVEC, as assessed by inhibition of binding with antibody to each protein. Lung microvascular endothelial cells had approximately half the number of HK binding sites as HUVEC while dermal micro vascular endothelial cells and astrocytes had only 8-10% of the sites/cell. The affinity of binding to the microvascular endothelial cells was greater than HUVEC, the affinity of binding to astrocytes was considerably less, nevertheless binding to each cell type involves gC1qR, cytokeratin 1 and u-PAR to varying degrees. We also demonstrate, for the first time, that factor XII binds to all of these cell types in a saturable and Zn(+2) dependent manner. Given that factor XII accelerates the interactions among cell surfaces and proteins of the contact activation cascade to generate bradykinin, binding of factor XII (and the prekallikrein-HK complex) may serve as a mechanism by which these proteins are concentrated locally to facilitate their interactions.
Department of Medicine, Division of Pulmonary and Critical Care, Allergy and Clinical Immunology, Medical University of South Carolina, P.O. Box 250623, 96 Jonathan Lucas Street, Charleston, SC 29425, USA.
Lawrence P
LP
Fernando
Snehlatha
S
Natesan
Kusumam
K
Joseph
Allen P
AP
Kaplan
D001253:Astrocytes / Q000378:metabolism*; D002352:Carrier Proteins; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005174:Factor XII / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors; D007633:Keratins / Q000378:metabolism; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism*; D008168:Lung / Q000098:blood supply; D008562:Membrane Glycoproteins; D008833:Microcirculation; D024101:Mitochondrial Proteins; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism; D011951:Receptors, Complement / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D012867:Skin / Q000098:blood supply; D014471:Umbilical Veins
C087299:C1QBP protein, human; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D019679:Kininogen, High-Molecular-Weight; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D011951:Receptors, Complement; D055293:Receptors, Urokinase Plasminogen Activator; C032402:complement 1q receptor; D007633:Keratins; D005174:Factor XII