The recognition sites of the integrins alpha1beta1 and alpha2beta1 within collagen IV are protected against gelatinase A attack in the native protein.
The susceptibility of three different solubilized forms of type IV collagen to gelatinase A cleavage and the concomitant effects on cell and integrin binding have been assessed. Dithiothreitol-solubilized Engelbreth-Holm Swarm (EHS) type IV collagen with disrupted intramolecular disulfide bonds in the CB3[IV] region was cleaved N-terminally to the CB3[IV] region into the two characteristic 100-300-nm fragments at 30 degrees C and was totally degraded at 37 degrees C. This was reflected in the partial or total loss of the alpha1beta1 and alpha2beta1 integrin binding sites within this region. The ability of gelatinase A to cleave EHS type IV collagen preparations with intact interchain disulfide bonds in CB3[IV] only occurred at higher temperatures. Furthermore, no effect on binding of cells or isolated integrins to the gelatinase-treated collagen could be detected after treatment at 37 degrees C. Dimeric collagen IV of human placenta with intact disulfide bonds in the CB3[IV] region was not degraded at all by gelatinase A at 37 degrees C.
Eble
J A
JA
Max-Planck-Institut für Biochemie, D-82152 Martinsried bei München, Germany.
Ries
A
A
Lichy
A
A
Mann
K
K
Stanton
H
H
Gavrilovic
J
J
Murphy
G
G
Kühn
K
K
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry*; D001665:Binding Sites; D002448:Cell Adhesion; D003094:Collagen / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D018093:Gelatinases / Q000378:metabolism*; D006801:Humans; D039423:Integrin alpha1; D039421:Integrin alpha2; D019012:Integrin beta1 / Q000737:chemistry*; D046911:Macromolecular Substances; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D051381:Rats; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D004220:Disulfides; D039423:Integrin alpha1; D039421:Integrin alpha2; D019012:Integrin beta1; D046911:Macromolecular Substances; D010446:Peptide Fragments; D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2