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https://hdl.handle.net/2440/5661
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Type: | Journal article |
Title: | Protein interaction studies of MAGP-1 with tropoelastin and fibrillin-1 |
Author: | Jensen, S. Reinhardt, D. Gibson, M. Weiss, A. |
Citation: | Journal of Biological Chemistry, 2001; 276(43):39661-39666 |
Publisher: | Amer Soc Biochemistry Molecular Biology Inc |
Issue Date: | 2001 |
ISSN: | 0021-9258 1083-351X |
Statement of Responsibility: | Sacha A. Jensen, Dieter P. Reinhardt, Mark A. Gibson and Anthony S. Weiss |
Abstract: | Elastic fibers consist primarily of an amorphous elastin core associated with microfibrils, 10-12 nm in diameter, containing fibrillins and microfibril-associated glycoproteins (MAGPs). To investigate the interaction of MAGP-1 with tropoelastin and fibrillin-1, we expressed human MAGP-1 as a T7-tag fusion protein in Escherichia coli. Refolding of the purified protein produced a soluble form of MAGP-1 that displayed saturable binding to tropoelastin. Fragments of tropoelastin corresponding to the N-terminal, C-terminal, and central regions of the molecule were used to characterize the MAGP-1 binding site. Cleavage of tropoelastin with kallikrein, which cleaves after Arg515 in the central region of the molecule, disrupted the interaction, suggesting that the separated N- and C-terminal fragments were insufficient to determine MAGP-1 binding to intact tropoelastin. In addition, no evidence of an interaction was observed between MAGP-1 and a tropoelastin construct consisting of domains 17-27 that brackets the kallikrein cleavage site, suggesting a complex mechanism of interaction between the two molecules. Binding of MAGP-1 was also tested with overlapping recombinant fibrillin-1 fragments. MAGP-1 bound to a region at the N terminus of fibrillin-1 in a calcium-dependent manner. In summary, these results suggest a model for the interaction of elastin with the microfibrillar scaffold. |
Keywords: | Humans Elastin Microfilament Proteins Oligopeptides Peptide Fragments Contractile Proteins Tropoelastin Recombinant Fusion Proteins Extracellular Matrix Proteins Protein Binding Fibrillin-1 Fibrillins RNA Splicing Factors |
Description: | © American Society for Biochemistry and Molecular Biology |
DOI: | 10.1074/jbc.M104533200 |
Published version: | http://www.jbc.org/cgi/content/abstract/276/43/39661 |
Appears in Collections: | Aurora harvest Pathology publications |
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