Identification of structural determinants controlling human and mouse stromelysin-3 proteolytic activities.
Matrix metalloproteinases (matrixins) constitute a group of extracellular proteinases belonging to the metzincin superfamily. They are involved in both physiological and pathological tissue remodeling processes, including those associated with cancer progression. Stromelysin-3, which is expressed in most invasive human carcinomas, is a matrix metalloproteinase with unusual functional properties. In particular, its mature form does not cleave any of the major extracellular matrix components. To define critical structural determinants involved in controlling stromelysin-3 proteolytic activity, we have used site-directed mutagenesis. We show that the deletion of at least 175 C-terminal amino-acids is sufficient to endow mouse stromelysin-3 with activities against casein, laminin, and type IV collagen. In the case of the human enzyme, however, a further and single Ala-235-->Pro substitution is necessary to observe similar activities. Ala-235, which characterizes human stromelysin-3 among matrixins, is located immediately after the C terminus of the "Met-turn," which forms a hydrophobic basis for the catalytic zinc atom in the metzincin family. We conclude that human stromelysin-3 has gained specific functional properties during evolution by amino acid substitution in the catalytic zinc environment, and that it represents an attractive target for specific inhibitors that may be used to prevent cancer progression.
Noël
A
A
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, BP163, Illkirch, CU de Strasbourg, France.
Santavicca
M
M
Stoll
I
I
L'Hoir
C
C
Staub
A
A
Murphy
G
G
Rio
M C
MC
Basset
P
P
D000409:Alanine; D000818:Animals; D001483:Base Sequence; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D003094:Collagen / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary; D006801:Humans; D007700:Kinetics; D007797:Laminin / Q000378:metabolism; D053507:Matrix Metalloproteinase 11; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011392:Proline; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity
D002364:Caseins; D017931:DNA Primers; D018076:DNA, Complementary; D007797:Laminin; D011994:Recombinant Proteins; D003094:Collagen; D011392:Proline; D053507:Matrix Metalloproteinase 11; D008666:Metalloendopeptidases; D000409:Alanine