title: Enzymatically crosslinked collagen-mimetic dendrimers that promote integrin-targeted cell adhesion.
abstract: Collagen is made up of a diverse family of the extracellular matrices, most of which are generally found crosslinked in vivo. To more closely mimic the biological function of collagen, this work focuses on establishing a molecular strategy to engineer a functional biomimetic collagen that exhibits stable collagen-like triple-helical conformation with cell-binding activity, in addition to an enzyme-mediated crosslinking by tissue transglutaminase (tTGase). A novel sequence spanning residues 2800-2807 of human fibrillin-1 (EDGFFKI) was first identified as an amine donor substrate for tTGase, using a previously characterized APQQEA derived from human osteonectin as an amine acceptor probe. Subsequently, collagen-mimetic peptides (CMPs) supplemented with a cell-binding sequence (GFOGER) and the identified EDGFFKI and APQQEA substrate sequences were conjugated onto a generation 2 poly(amidoamine) dendrimer, resulting in a crosslinkable collagen-mimetic dendrimer, denoted as CMD-K and CMD-Q, respectively. Both CMD-K and CMD-Q exhibited enhanced triple-helical stability and supported cell adhesion in an integrin-specific manner. Finally, tTGase-mediated crosslinking between CMD-K and CMD-Q resulted in a supramolecular structure that exhibited stable collagen-like triple-helical conformation and improved cellular recognition. The results show that the triple-helical structure is important in preserving the GFOGER cell-binding site while the tTGase-mediated protein crosslinking may also be crucial for the recognition by cell surface integrin receptors.
authors.affiliation: Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore 119077, Singapore. st.khew@nus.edu.sg
authors.forename: Shih Tak
authors.initials: ST
authors.lastname: Khew
authors.forename: Qing Jun
authors.initials: QJ
authors.lastname: Yang
authors.forename: Yen Wah
authors.initials: YW
authors.lastname: Tong
mesh_terms: D000595:Amino Acid Sequence; D002448:Cell Adhesion; D003094:Collagen / Q000737:chemistry*; D050091:Dendrimers; D000071838:Fibrillin-1; D000071837:Fibrillins; D006801:Humans; D016023:Integrins / Q000737:chemistry*; D008840:Microfilament Proteins / Q000737:chemistry; D018716:Molecular Mimicry; D054392:Adipokines
chemical_list: D003094:Collagen; D050091:Dendrimers; D000071838:Fibrillin-1; D000071837:Fibrillins; D016023:Integrins; D008840:Microfilament Proteins; C000605722:FBN1 protein, human; D054392:Adipokines