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J. Biol. Chem., Vol. 277, Issue 38, 35044-35049, September 20, 2002
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From the Elastic fibers are composed of the protein
elastin and a network of 10-12 nm microfibrils. The microfibrillar
proteins include, among others, the fibrillins and
microfibril-associated glycoproteins-1 and -2 (MAGP-1 and MAGP-2).
Little is known about how microfibrillar proteins interact to support
fiber assembly. We used the C-terminal half of MAGP-2 in a yeast
two-hybrid library screen to identify relevant ligands. Six of 13 positive clones encoded known microfibrillar proteins, including
fibrillin-1 and -2. Deletion analysis of partial fibrillin-1 and -2 clones revealed a calcium-binding epidermal growth factor
repeat-containing region near the C terminus responsible for binding.
This region is distinct from the region of fibrillin-1 reported by
others to bind MAGP-1. The MAGP-2 bait was unable to interact
productively with other epidermal growth factor repeats in fibrillin-1,
demonstrating specificity of the interaction. Deletion analysis of the
MAGP-2 bait demonstrated that binding occurred in a core region
containing 48% identity and 7 conserved cysteine residues with MAGP-1.
Immunoprecipitation of MAGP-2 from transfected COS-7 cells
resulted in the coprecipitation of fibrillin. These results demonstrate
that MAGP-2 specifically interacts with fibrillin-1 and -2 and suggest
that MAGP-2 may help regulate microfibrillar assembly. The results also
demonstrate the utility of the yeast two-hybrid system to study
protein-protein interactions of the extracellular matrix.
Microfibril-associated Glycoprotein-2 Interacts with Fibrillin-1
and Fibrillin-2 Suggesting a Role for MAGP-2 in Elastic Fiber
Assembly*
,¶
Division of Pulmonary and Critical Care
Medicine, Department of Internal Medicine, Barnes-Jewish Hospital at
Washington University School of Medicine, St. Louis, Missouri 63110 and the § Wellcome Trust Centre for Cell-Matrix Research,
University of Manchester,
Manchester M13 9PT, United Kingdom
*
This work was supported by the Barnes-Jewish Hospital
Foundation (to J. M. S.), National Institutes of Health Grant
HL67353, an American Lung Association Career Investigator award (to
J. M. S.), and the Alan A. and Edith L. Wolff Charitable Trust.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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