Role of ADAM-9 disintegrin-cysteine-rich domains in human keratinocyte migration.
ADAM-9 belongs to a family of transmembrane, disintegrin-containing metalloproteinases involved in protein ectodomain shedding and cell-cell and cell-matrix interactions. The aim of this study was to analyze the expression of ADAM-9 in skin and to assess the role of this proteolytic/adhesive protein in skin physiology. In normal skin, ADAM-9 expression was detected in both the epidermis and dermis and in vitro in keratinocytes and fibroblasts. Here we report that ADAM-9 functions as a cell adhesion molecule via its disintegrin-cysteine-rich domain. Using solid phase binding assays and antibody inhibition experiments, we demonstrated that the recombinant disintegrin-cysteine-rich domain of ADAM-9 specifically interacts with the beta1 integrin subunit on keratinocytes. This was corroborated by co-immunoprecipitation. In addition, engagement of integrin receptors by the disintegrin-cysteine-rich domain resulted in ERK phosphorylation and increased MMP-9 synthesis. Treatment with the ERK inhibitor PD98059 inhibited MMP-9 induction. Furthermore, the presence of the soluble disintegrin-cysteine-rich domain did not interfere with cell migration on different substrates. However, keratinocytes adhering to the immobilized disintegrin-cysteine-rich domain showed increased motility, which was partially due to the induction of MMP-9 secretion. In summary, our results indicate that the ADAM-9 adhesive domain plays a role in regulating the motility of cells by interaction with beta1 integrins and modulates MMP synthesis.
Zigrino
Paola
P
Department of Dermatology and Center for Molecular Medicine, Kerpener Strasse 62, University of Cologne, 50937 Cologne, Germany. Paola.Zigrino@uni-koeln.de
Steiger
Julia
J
Fox
Jay W
JW
Löffek
Stefanie
S
Schild
Alexander
A
Nischt
Roswitha
R
Mauch
Cornelia
C
D051722:ADAM Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D020405:Dermis / Q000166:cytology / Q000201:enzymology; D019483:Disintegrins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D000078404:Epidermal Cells; D004817:Epidermis / Q000201:enzymology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology*; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000502:physiology; D012879:Skin Physiological Phenomena
D015815:Cell Adhesion Molecules; D019483:Disintegrins; D005419:Flavonoids; D019012:Integrin beta1; D008565:Membrane Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D051722:ADAM Proteins; C098339:ADAM9 protein, human; D020780:Matrix Metalloproteinase 9; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one