Screening for novel cell adhesive regions in bovine Achilles tendon collagen peptides.
Collagen, a major structural protein of the ECM, is known for its high cell adherence capacity. This study was conducted to identify regions in collagen that harbour such bioactivity. Collagen from tendon was hydrolysed and the peptides fractionated using ion-exchange chromatography (IEC). Isolated peptide fractions were coated onto disposable dishes and screened for cell adherence and proliferative abilities. Active IEC fractions were further purified by chromatography, and two peptides, C2 and E1 with cell adhesion ability, were isolated. A cell adhesion assay done with different amounts of C2 coated onto disposable dishes revealed the maximum adhesion to be 94.6%, compared with 80% for collagen coated dishes and an optimum peptide coating density of 0.507 nmoles per cm(2) area of the dish. Growth of cells on C2, collagen, and E1 revealed a similar pattern and a reduction in the doubling time compared with cells grown on uncoated dishes. C2 had a mass of 2.046 kDa with 22 residues, and sequence analysis revealed a higher percentage occurrence of hydrophilic residues compared with other regions in collagen. Docking studies revealed GDDGEA in C2 as the probable site of interaction with integrins α2β1 and α1β1, and stability studies proved C2 to be mostly protease-resistant.
School of Bio Science and Technology, VIT University, Vellore 632014, Tamil Nadu, India.
Pradipta
P
Banerjee
Alka
A
Mehta
C
C
Shanthi
D000125:Achilles Tendon / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects*; D002522:Chlorocebus aethiops; D024042:Collagen Type I / Q000737:chemistry*; D003198:Computer Simulation; D006367:HeLa Cells; D006801:Humans; D016023:Integrins / Q000737:chemistry; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D014709:Vero Cells
D024042:Collagen Type I; D016023:Integrins; D010455:Peptides