Identification and validation of a novel cell-recognition site (KNEED) on the 8th type III domain of fibronectin.
Interactions between cell-surface integrins and extracellular matrix proteins underlie a versatile recognition system providing cells with anchorage, traction for migration or matrix remodeling, as well as signals for polarity, differentiation and growth. Short peptide sequences of fibronectin (FN), most notably RGD found on a loop in the 10th type III domain, are effective in promoting cell adhesion when immobilized to a biomaterial scaffold. Additional sequences (e.g. PHSRN) have been shown to act synergistically to enhance cell adhesion and other cellular processes. Using bioinformatics, we identified a candidate cell-binding peptide sequence, KNEED, located on the loop region of the 8th domain of FN that from in vitro studies appears to participate in cell attachment and spreading. Computational analysis revealed that KNEED exhibits both high solvent accessibility and sequence conservation values across FN sequences from seven species. We demonstrate the importance of the KNEED sequence using a solution-phase competitive inhibition assay utilizing soluble peptides. Results indicate that the presence of soluble KNEED peptides inhibits the attachment and spreading of 3T3 balb/c fibroblasts on FN-coated surfaces in a concentration-dependent manner. As more sequence and crystallographic data become available, computational approaches may aid in the identification of new targets for applications where biorecognition plays a key role.
Wong
Joyce Y
JY
Department of Biomedical Engineering, Boston University, MA 02215, USA. jywong@bu.edu
Weng
Zhiping
Z
Moll
Sarah
S
Kim
Sooyoung
S
Brown
Christopher T
CT
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001672:Biocompatible Materials; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D002465:Cell Movement; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008858:Microscopy, Phase-Contrast; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
D001672:Biocompatible Materials; D005353:Fibronectins