The use of collagen-based model peptides to investigate platelet-reactive sequences in collagen

Barnes, Michael J.; Knight, Christopher G.; Farndale, Richard W.

doi:10.1002/(sici)1097-0282(1996)40:4<383::aid-bip4>3.0.co;2-s

Cited by 33 publications

(25 citation statements)

References 75 publications

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“…Several important clues to the understanding of the interaction of ␣ 2 ␤ 1 integrin with collagen arose from the development of triple helical synthetic peptides containing designated recognition sequences (6). From these studies, a GFOGER sequence was identified as a crucial, high affinity ␣ 1 ␤ 1 and ␣ 2 ␤ 1 integrin binding site in collagen I (7).…”

mentioning

confidence: 99%

Prolyl Hydroxylation of Collagen Type I Is Required for Efficient Binding to Integrin α1β1 and Platelet Glycoprotein VI but Not to α2β1

Perret

Eble

Siljander

et al. 2003

Journal of Biological Chemistry

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Collagen is a potent adhesive substrate for cells, an event essentially mediated by the integrins ␣ 1 ␤ 1 and ␣ 2 ␤ 1 . Collagen fibrils also bind to the integrin ␣ 2 ␤ 1 and the platelet receptor glycoprotein VI to activate and aggregate platelets. The distinct triple helical recognition motifs for these receptors, GXOGER and (GPO) n , respectively, all contain hydroxyproline. Using unhydroxylated collagen I produced in transgenic plants, we investigated the role of hydroxyproline in the receptorbinding properties of collagen. We show that ␣ 2 ␤ 1 but not ␣ 1 ␤ 1 mediates cell adhesion to unhydroxylated collagen. Soluble recombinant ␣ 1 ␤ 1 binding to unhydroxylated collagen is considerably reduced compared with bovine collagens, but binding can be restored by prolyl hydroxylation of recombinant collagen. We also show that platelets use ␣ 2 ␤ 1 to adhere to the unhydroxylated recombinant molecules, but the adhesion is weaker than on fully hydroxylated collagen, and the unhydroxylated collagen fibrils fail to aggregate platelets. Prolyl hydroxylation is thus required for binding of collagen to platelet glycoprotein VI and to cells by ␣ 1 ␤ 1 . These observations give new insights into the molecular basis of collagen-receptor interactions and offer new selective applications for the recombinant unhydroxylated collagen I.The collagens include the most abundant proteins in mammalian tissues providing a scaffold and framework for extracellular matrix assembly. In addition, they can modulate cell behavior through interactions with cellular receptors. All members of the collagen family are built up of three chains and contain at least one triple helix domain formed by repeating GXY sequences (where X is often proline (P) and Y is often hydroxyproline (O)). Such molecules associate to form complex structures, of which the fibril-forming collagens constitute the most abundant matrix component (1). The higher order organization of the collagens, collagen I being the best example, proved crucial to the triggering of specific cell responses; native triple helical structure is essential for cell and platelet adhesion, and the fibrillar structure is required for platelet activation and aggregation (2).The cell surface receptors ␣ 1 ␤ 1 and ␣ 2 ␤ 1 of the integrin family are principally collagen receptors, although they can interact with other extracellular matrix components (e.g. laminins). The specificity of the more recently discovered collagen binding integrins, ␣ 10 ␤ 1 and ␣ 11 ␤ 1 , remains to be determined. Different receptor families have also been identified as containing collagen receptors, such as the membrane-spanning proteoglycans, syndecans (3), and the recently described tyrosine kinase receptors DDR1 and DDR2 (4). In platelets, the recently described glycoprotein VI is a key signaling collagen receptor and is an immunoglobulinn superfamily receptor (5).Several important clues to the understanding of the interaction of ␣ 2 ␤ 1 integrin with collagen arose from the development of triple helical synthetic peptid...

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confidence: 99%

Prolyl Hydroxylation of Collagen Type I Is Required for Efficient Binding to Integrin α1β1 and Platelet Glycoprotein VI but Not to α2β1

Perret

Eble

Siljander

et al. 2003

Journal of Biological Chemistry

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“…On the basis of inhibition of platelet adhesion to this collagen by short, linear (non-triple-helical) peptides, an ␣ 2 ␤ 1 -binding site has been assigned to the sequence DGEA, which corresponds to residues 435-438 of the ␣1(I) chain and is found in the CNBr-derived fragment ␣1(I)CB3 (10). 2 However, others have observed no inhibition of ␣ 2 ␤ 1 -mediated cell adhesion to collagen by DGEAcontaining peptides (9,(11)(12)(13)(14)(15). Platelet adhesion to ␣1(I)CB3 is ␣ 2 ␤ 1 -dependent (9,16), but the fragment exhibits little platelet aggregatory activity (17).…”

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confidence: 99%

“…Here we describe studies on the ability of these peptides to support platelet adhesion and activation, and the adhesion of HT 1080 cells, which also express the integrin ␣ 2 ␤ 1 . A preliminary account of some of this work has been given (14).…”

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confidence: 99%

The Platelet Reactivity of Synthetic Peptides Based on the Collagen III Fragment α1(III)CB4

Morton¹,

Peachey²,

Knight³

et al. 1997

Journal of Biological Chemistry

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The platelet-reactive collagen III-derived fragment ␣1(III)CB4 has been synthesized as seven overlapping peptides, each as a homotrimeric triple-helical species covalently linked at the C terminus. Additional Gly-ProHyp triplets were introduced at each end of the peptide sequence to ensure a stable triple-helical conformation at 20°C, the temperature at which cell reactivity was measured. A Cys-containing triplet was included at each end to allow intermolecular cross-linking. All seven peptides in triple-helical, cross-linked form were able to cause platelet aggregation. Peptide 6, the most reactive species, was more aggregatory than collagen fibers. Platelet adhesion occurred to all peptides immobilized on plastic in monomeric form. Adhesion was integrin ␣ 2 ␤ 1 -independent except in the case of peptide 6, adhesion to which was partially reduced by anti-integrin ␣ 2 ␤ 1 monoclonal antibodies. The presence of an ␣ 2 ␤ 1 recognition site in peptide 6 was confirmed using HT 1080 cells, which express ␣ 2 ␤ 1 as their major or sole collagen receptor. HT 1080 adhesion to both peptide 6 and collagen was strongly inhibited by anti-integrin ␣ 2 ␤ 1 monoclonal antibodies. These cells did not adhere to any of the other peptides. Comparison of the structure of peptide 6 with that of adjacent peptides indicates that the sequence Gly-Gly-Pro-Hyp-Gly-Pro-Arg, residues 522-528 of the collagen ␣1(III) chain, represents the minimum structure required for the recognition of ␣ 2 ␤ 1 . Our findings support the view that the collagen triple helix possesses an intrinsic platelet reactivity that can be expressed independently of integrin ␣ 2 ␤ 1 and the precise level of which is governed by the exact nature of the primary sequence. Sequences such as those recognizing ␣ 2 ␤ 1 may potentiate the activity, whereas others may have the opposite effect.Cell interaction with collagens is mediated by specific cellsurface receptors, important among which are the integrins ␣ 1 ␤ 1 and ␣ 2 ␤ 1 (1). Integrins recognize discrete amino acid sequences, the best known of which, perhaps, is the RGD 1 sequence, which serves as a cell-binding site in several matrix proteins and is recognized by a number of integrins, including the ␤ 1 -integrins ␣

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“…Our flow cytometric results clearly showed that ADP release from the platelets is an important part of the response to stronger stimuli such as the collagen-related peptide. This peptide has been shown to increase intracellular calcium levels, activate protein kinase C and phosphorylate phospholipase C2 even in the presence of apyrase in the medium, but it could not decrease the adenylate cyclase activity [28]. This could explain why blocking the P2Y 12 receptor was more effective in inhibiting platelet fibrinogen binding after CRP stimulation.…”

Section: Discussionmentioning

confidence: 59%

Effects of inhibition of P2Y1 and P2Y12 on whole blood clotting, coagulum elasticity and fibrinolysis resistance studied with free oscillation rheometry

Ramström

Rånby

Lindahl³

2003

Thrombosis Research

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The use of collagen-based model peptides to investigate platelet-reactive sequences in collagen

Cited by 33 publications

References 75 publications

Prolyl Hydroxylation of Collagen Type I Is Required for Efficient Binding to Integrin α1β1 and Platelet Glycoprotein VI but Not to α2β1

Prolyl Hydroxylation of Collagen Type I Is Required for Efficient Binding to Integrin α1β1 and Platelet Glycoprotein VI but Not to α2β1

The Platelet Reactivity of Synthetic Peptides Based on the Collagen III Fragment α1(III)CB4

Effects of inhibition of P2Y1 and P2Y12 on whole blood clotting, coagulum elasticity and fibrinolysis resistance studied with free oscillation rheometry

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