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Engineering autoactivating forms of matrix metalloproteinase-9 and expression of the active enzyme in cultured cells
Katherine E Fisher1, Qing Fei, Ellen R Laird
1Department of CNS Discovery, Global Research and Development, Groton Laboratories, Pfizer, Inc., Groton, Connecticut 06340-5146, USA. katerine_e_fisher@groton.pfizer.com
Abstract:
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.
Insights
Matrix metalloproteinases (MMPs) are implicated in central nervous system disorders. Researchers engineered autoactive MMP-9 forms to study its role in diseases like Alzheimer's and stroke.
Area of Science:
- Neuroscience
- Biochemistry
- Enzymology
Background:
- Matrix metalloproteinases (MMPs), particularly MMP-9 (gelatinase B) and MMP-2 (gelatinase A), are increasingly linked to the pathogenesis of neurological disorders.
- Elevated MMP expression is observed in conditions such as Alzheimer's disease, stroke, multiple sclerosis, and amyotrophic lateral sclerosis, suggesting dysregulation.
- This aberrant regulation may lead to inappropriate MMP activity, contributing to disease progression.
Purpose of the Study:
- To investigate the impact of elevated active matrix metalloproteinase-9 (MMP-9) levels within the central nervous system.
- To create tools for studying the consequences of dysregulated MMP-9 activity in neurological diseases.
Main Methods:
- Designed mutant forms of matrix metalloproteinase-9 (MMP-9) to enable autocatalytic removal of the pro domain, resulting in active enzyme.
- Modified specific residues within the cysteine switch autoinhibitor region of the MMP-9 propeptide.
- Developed stable cell lines and transgenic mouse models expressing autoactive G100L and D103N forms of human MMP-9.
Main Results:
- Successfully generated mutant MMP-9 enzymes capable of autocatalytic activation.
- Established stable cell lines and transgenic mice expressing these autoactive MMP-9 variants.
- These models allow for the study of increased active MMP-9 levels in vivo and in vitro.
Conclusions:
- The developed autoactive MMP-9 models provide a valuable resource for elucidating the role of MMP-9 dysregulation in central nervous system disorders.
- Further research using these models can help understand the specific mechanisms by which MMP-9 contributes to neurodegeneration and neurological disease progression.
- This work facilitates the evaluation of therapeutic strategies targeting MMP-9 activity in CNS diseases.
