 | Bethany Walton Ramapo College of New Jersey BiochemistryMentor(s)Barbara Brodsky, Ph.D. Teresita Silva Department of Biochemistry UMDNJ-Robert Wood Johnson Medical School |
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| Characterization of the Properties of Collagen-Like Domains from Bacterial Collagen | ||
| Collagen is an important structural triple-helical protein found in animals and also in certain bacteria. Studying the characteristics of the bacterial collagen extends general understanding of the collagen triple helix. The specific aim of this project was to characterize the structural properties of collagen-like domains from four bacterial proteins: Methylobacterium, Clostridium perfringens, Solibacter usitatus, and Rhodopseudomonas palustris. Experimental methods were used to study their triple helix structures. Circular dichroism (CD) spectra were obtained to test for the presence of the triple helix structure of the domains. The spectra for Methylobacterium, C. perfringens and S. usitatus had features of a typical triple helix. The ratio of the magnitude of the positive 225 nm and negative 198 nm peaks was used to assess if the protein was a complete triple helix. The results showed that C. perfringens and S. usitatus have a more triple helical conformation than Methylobacterium. Dynamic light scattering (DLS) was done to check for aggregation and determine the hydrodynamic radius of the domains. As expected, the hydrodynamic radius (Rh) obtained from DLS increased as the length of the triple helix domain increased except for Methylobacterium, which showed a higher Rh value which may suggest aggregation. Computational methods were used to analyze the sequences of the bacterial collagen. The collagen-like domains of the different proteins were categorized by the nature of their residues as charged, polar, proline, small or hydrophobic and showed varying amino acid compositions. The domains were analyzed for repeating sequences and for hydrophobic periodicity. They differed in the number and length of repeating amino acid sequence and periodicity. Characterization of the triple helix and domain sequence may lead to the development of recombinant bacterial collagen that can be used as models for collagen diseases and have biomaterial applications. |