Defining the domains of type I collagen involved in heparin- binding and endothelial tube formation.
Cell surface heparan sulfate proteoglycan (HSPG) interactions with type I collagen may be a ubiquitous cell adhesion mechanism. However, the HSPG binding sites on type I collagen are unknown. Previously we mapped heparin binding to the vicinity of the type I collagen N terminus by electron microscopy. The present study has identified type I collagen sequences used for heparin binding and endothelial cell-collagen interactions. Using affinity coelectrophoresis, we found heparin to bind as follows: to type I collagen with high affinity (Kd approximately 150 nM); triple-helical peptides (THPs) including the basic N-terminal sequence alpha1(I)87-92, KGHRGF, with intermediate affinities (Kd approximately 2 microM); and THPs including other collagenous sequences, or single-stranded sequences, negligibly (Kd >> 10 microM). Thus, heparin-type I collagen binding likely relies on an N-terminal basic triple-helical domain represented once within each monomer, and at multiple sites within fibrils. We next defined the features of type I collagen necessary for angiogenesis in a system in which type I collagen and heparin rapidly induce endothelial tube formation in vitro. When peptides, denatured or monomeric type I collagen, or type V collagen was substituted for type I collagen, no tubes formed. However, when peptides and type I collagen were tested together, only the most heparin-avid THPs inhibited tube formation, likely by influencing cell interactions with collagen-heparin complexes. Thus, induction of endothelial tube morphogenesis by type I collagen may depend upon its triple-helical and fibrillar conformations and on the N-terminal heparin-binding site identified here.
Sweeney
S M
SM
Department of Medicine and the Cardeza Foundation for Hematologic Research, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107, USA.
Guy
C A
CA
Fields
G B
GB
San Antonio
J D
JD
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D013499:Surface Properties
D006493:Heparin; D003094:Collagen