Homotypic fibrillin-1 interactions in microfibril assembly.
We have defined the homotypic interactions of fibrillin-1 to obtain new insights into microfibril assembly. Dose-dependent saturable high affinity binding was demonstrated between N-terminal fragments, between furin processed C-terminal fragments, and between these N- and C-terminal fragments. The N terminus also interacted with a downstream fragment. A post-furin cleavage site C-terminal sequence also interacted with the N terminus, with itself and with the furin-processed fragment. No other homotypic fibrillin-1 interactions were detected. Some terminal homotypic interactions were inhibited by other terminal sequences, and were strongly calcium-dependent. Treatment of an N-terminal fragment with N-ethylmaleimide reduced homotypic binding. Microfibril-associated glycoprotein-1 inhibited N- to C-terminal interactions but not homotypic N-terminal interactions. These fibrillin-1 interactions are likely to regulate pericellular fibrillin-1 microfibril assembly.
Marson
Andrew
A
Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
Rock
Matthew J
MJ
Cain
Stuart A
SA
Freeman
Lyle J
LJ
Morgan
Amanda
A
Mellody
Kieran
K
Shuttleworth
C Adrian
CA
Baldock
Clair
C
Kielty
Cay M
CM
D020816:Amino Acid Motifs; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D003285:Contractile Proteins / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005033:Ethylmaleimide / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D000071838:Fibrillin-1; D000071837:Fibrillins; D045683:Furin / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008024:Ligands; D016014:Linear Models; D020894:Microfibrils / Q000378:metabolism*; D008840:Microfilament Proteins / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D000072260:RNA Splicing Factors; D011994:Recombinant Proteins / Q000737:chemistry; D054392:Adipokines
D002118:Calcium; D003285:Contractile Proteins; D005033:Ethylmaleimide; D016326:Extracellular Matrix Proteins; D000071838:Fibrillin-1; D000071837:Fibrillins; D045683:Furin; D008024:Ligands; D008840:Microfilament Proteins; D000072260:RNA Splicing Factors; D011994:Recombinant Proteins; C000605722:FBN1 protein, human; D054392:Adipokines; C016598:microfibrillar protein