Engineering autoactivating forms of matrix metalloproteinase-9 and expression of the active enzyme in cultured cells and transgenic mouse brain.
Matrix metalloproteinases (MMPs) are hypothesized to play an important role in the pathogenesis of several central nervous system disorders. Increased levels of expression of MMP-9 (gelatinase B) and MMP-2 (gelatinase A) have been observed in Alzheimer's disease, stroke, multiple sclerosis, and amyotrophic lateral sclerosis. This suggests an aberrant regulation of MMPs that could lead to inappropriate expression of MMP activity. To allow us to evaluate the effect of increased levels of active MMP-9 in the central nervous system, mutant forms of the enzyme were designed to autocatalytically remove the pro domain, yielding active enzyme. This was accomplished by modifying residues in the cysteine switch autoinhibitor region of the propeptide. Stable cell lines and transgenic mice that express G100L and D103N autoactive forms of human MMP-9 were developed to study the role of dysregulation of MMP-9 in disease.
Fisher
Katherine E
KE
Department of CNS Discovery, Global Research and Development, Groton Laboratories, Pfizer, Inc., Groton, Connecticut 06340-5146, USA. katerine_e_fisher@groton.pfizer.com
Fei
Qing
Q
Laird
Ellen R
ER
Stock
Jeffrey L
JL
Allen
Melanie R
MR
Sahagan
Barbara G
BG
Strick
Christine A
CA
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D001921:Brain / Q000201:enzymology*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017931:DNA Primers; D006624:Hippocampus / Q000201:enzymology; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017931:DNA Primers; D016212:Transforming Growth Factor beta; D020780:Matrix Metalloproteinase 9