The adhesion and migration of cells within triple-helical regions of native collagens is an essential process in both normal and pathological conditions. Five specific sequences (9-15 residues long) within triple- helical regions of native collagens have been identified as adhesion and migration sites for a variety of cell types. Prior studies have examined cell adhesion to single-stranded peptide models of these sequences; thus, the relative influences of collagen primary and secondary structure on these cell interactions has not been ascertained. We will chemically synthesize triple-helical and non-triple peptides from types I-IV collagen and use well documented assays to study the relative importance of collagen primary and secondary structure by comparing triple-helical and non-triple-helical peptides for their ability to promote cell adhesion and migration. We will determine if cell adhesion is mediated by the same cell-surface integrin at all five sequences or if different integrins are responsible for adhesion to different sequences. The secondary and tertiary structure of single-stranded and triple-helical peptides will be investigated by circular dichroism and residue substitution or modification on triple-helical conformation, which in turn will correlated to changes in biological function. Using these peptides, we will determine how protein primary and secondary structure affects the affinity and specificity of cell adhesion.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Modified Research Career Development Award (K04)
Project #
5K04AR001929-04
Application #
2376607
Study Section
Biochemistry Study Section (BIO)
Project Start
1994-03-01
Project End
1999-02-28
Budget Start
1997-03-01
Budget End
1998-02-28
Support Year
4
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Pathology
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Lauer-Fields, Janelle L; Fields, Gregg B (2002) Triple-helical peptide analysis of collagenolytic protease activity. Biol Chem 383:1095-105
Malkar, Navdeep B; Lauer-Fields, Janelle L; Borgia, Jeffrey A et al. (2002) Modulation of triple-helical stability and subsequent melanoma cellular responses by single-site substitution of fluoroproline derivatives. Biochemistry 41:6054-64
Lauer-Fields, Janelle L; Juska, Darius; Fields, Gregg B (2002) Matrix metalloproteinases and collagen catabolism. Biopolymers 66:19-32
Lauer-Fields, J L; Broder, T; Sritharan, T et al. (2001) Kinetic analysis of matrix metalloproteinase activity using fluorogenic triple-helical substrates. Biochemistry 40:5795-803
Edwards, W B; Anderson, C J; Fields, G B et al. (2001) Evaluation of radiolabeled type IV collagen fragments as potential tumor imaging agents. Bioconjug Chem 12:1057-65
Fields, G B (2001) Using fluorogenic peptide substrates to assay matrix metalloproteinases. Methods Mol Biol 151:495-518
Forns, P; Lauer-Fields, J L; Gao, S et al. (2000) Induction of protein-like molecular architecture by monoalkyl hydrocarbon chains. Biopolymers 54:531-46
Lauer-Fields, J L; Nagase, H; Fields, G B (2000) Use of Edman degradation sequence analysis and matrix-assisted laser desorption/ionization mass spectrometry in designing substrates for matrix metalloproteinases. J Chromatogr A 890:117-25
Borgia, J A; Fields, G B (2000) Chemical synthesis of proteins. Trends Biotechnol 18:243-51
Lauer-Fields, J L; Tuzinski, K A; Shimokawa, K i et al. (2000) Hydrolysis of triple-helical collagen peptide models by matrix metalloproteinases. J Biol Chem 275:13282-90

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