Microfibril-associated glycoprotein binds to the carboxyl-terminal domain of tropoelastin and is a substrate for transglutaminase.
Microfibril-associated glycoprotein (MAGP) is an integral component of microfibrillar structures that play a critical role in the organization of elastic fibers in the extracellular matrix. To study possible molecular interactions between MAGP and other elastic fiber components, we have generated native MAGP using a baculovirus expression system and tested its ability to associate with tropoelastin and fibrillin. MAGP produced by SF9 cells underwent processing similar to the mammalian protein, including correct cleavage of the signal peptide and sulfation of tyrosine residues. When tested in solid-phase binding assays, native MAGP specifically bound to tropoelastin but not fibrillin-1. Binding to tropoelastin was divalent cation-independent and was completely blocked by reduction and alkylation of either protein. Antibody inhibition studies indicated that the carboxyl terminus of tropoelastin mediated its interaction with MAGP. In addition to binding to elastin, MAGP was also a substrate for transglutaminase, which might explain its propensity to form high molecular weight aggregates that cannot be dissociated with reduction or denaturation. Together, the results of this study provide new insights into the functional relationship between microfibrillar proteins and have important implications for understanding elastic fiber assembly.
Brown-Augsburger
P
P
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
Broekelmann
T
T
Mecham
L
L
Mercer
R
R
Gibson
M A
MA
Cleary
E G
EG
Abrams
W R
WR
Rosenbloom
J
J
Mecham
R P
RP
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D003285:Contractile Proteins; D018076:DNA, Complementary; D004549:Elastin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D000072260:RNA Splicing Factors; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera; D014162:Transfection; D011503:Transglutaminases / Q000378:metabolism*; D014333:Tropoelastin / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D003285:Contractile Proteins; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D010446:Peptide Fragments; D000072260:RNA Splicing Factors; D011994:Recombinant Proteins; D014333:Tropoelastin; C016598:microfibrillar protein; D004549:Elastin; D011503:Transglutaminases