A5021997935- Protease and Inhibitor Mechanisms
- Peptidase Inhibition and Analysis
- RNA modifications and cancer
- Biological Research and Disease Studies
- Cell Adhesion Molecules Research
- Epigenetics and DNA Methylation
- Signaling Pathways in Disease
- Immunotherapy and Immune Responses
- Cell death mechanisms and regulation
- Blood Coagulation and Thrombosis Mechanisms
- Bone and Dental Protein Studies
- Bacillus and Francisella bacterial research
- Bacterial Genetics and Biotechnology
- vaccines and immunoinformatics approaches
- RNA Interference and Gene Delivery
- Cancer Research and Treatments
- Pancreatic function and diabetes
- NF-κB Signaling Pathways
- Bioactive Compounds and Antitumor Agents
- Free Radicals and Antioxidants
- Diabetes and associated disorders
- Parasitic Diseases Research and Treatment
- Microbial Inactivation Methods
- Biochemical Acid Research Studies
- Molecular Biology Techniques and Applications
Oregon Health & Science University
2013-2021
Cancer Research Center
2003-2010
Sanford Burnham Prebys Medical Discovery Institute
2002-2010
Institute for Medical Research
2006-2008
Discovery Institute
2006
Torrey Pines Institute For Molecular Studies
2005
George Mason University
2005
Scripps Research Institute
2005
Recently, we have shown that membrane type 1 matrix metalloproteinase (MT1-MMP) exhibits integrin convertase activity. Similar to furin-like proprotein convertases, MT1-MMP directly processes a single chain precursor of αvintegrin subunit (pro-αv) into the heavy and light α-chains connected by disulfide bridge. To evaluate functionality MT1-MMP-processed integrins, examined breast carcinoma MCF7 cells co-expressing αvβ3 with either wild or mutant in variety migration adhesion tests. Specific...
Membrane type-1 matrix metalloproteinase (MT1-MMP) and α<sub>v</sub>β<sub>3</sub> integrin are both essential to cell invasion. Maturation of pro-α<sub>v</sub>chain (pro-α<sub>v</sub>) involves its cleavage by proprotein convertases (PC) form the disulfide-bonded 125-kDa heavy 25-kDa light α chains. Our report presents evidence an alternative pathway pro-α<sub>v</sub> processing involving MT1-MMP. In breast carcinoma MCF7 cells deficient in MT1-MMP, is processed a conventional furin-like PC,...
We demonstrate that the presentation of LRP and subsequent uptake its ligands by malignant cells are both strongly regulated MT1-MMP. Because is essential for clearance multiple ligands, these findings have important implications many pathophysiological processes including pericellular proteolysis in neoplastic as well fate soluble matrix-degrading proteases such MMP-2. MT1-MMP a key protease cell invasion physiological activator Cellular consists non-covalently associated 515-kDa...
Inhalation anthrax is a deadly disease for which there currently no effective treatment. Bacillus anthracis lethal factor (LF) metalloproteinase an integral component of the tripartite toxin that essential onset and progression anthrax. We report here on fragment-based approach allowed us to develop inhibitors LF. The small-molecule we have designed, synthesized, tested are highly potent selective against LF in both vitro tests cell-based assays. These do not affect prototype human...
Membrane-type 1 matrix metalloproteinase (MT1-MMP), a transmembrane proteinase with short cytoplasmic domain and an extracellular catalytic domain, controls variety of physiological pathological processes through the proteolytic degradation or proteins. MT1-MMP forms complex on cell membrane its protein inhibitor, tissue inhibitor metalloproteinases-2 (TIMP-2). Here we show that, in addition to proteolysis, TIMP-2 control proliferation migration non-proteolytic mechanism. binding induces...
Elevated expression of membrane type-1 matrix metalloproteinase (MT1-MMP) is closely associated with malignancies. There a consensus among scientists that cell surface-associated MT1-MMP key player in pericellular proteolytic events. Now we have identified an intracellular, hitherto unknown, function MT1-MMP. We demonstrated trafficked along the tubulin cytoskeleton. A fraction cellular accumulates centrosomal compartment. targets integral protein, pericentrin. Pericentrin known to be...
Mosquito-borne WNV (West Nile virus) is an emerging global threat. The NS3 proteinase, which essential for the proteolytic processing of viral polyprotein precursor, a promising drug target. We have isolated and biochemically characterized recombinant, highly active proteinase. determined that proteinase functions in manner distantly similar to furin cleaving peptide protein substrates. aprotinin D-arginine-based 9-12-mer peptides are potent inhibitors with K(i) values 26 nM 1 respectively....
Infiltration of T cells in breast tumors correlates with improved survival patients cancer, despite relatively few mutations these tumors. To determine if T-cell specificity can be harnessed to augment immunotherapies we sought identify the alpha-beta paired receptors (TCRs) tumor-infiltrating lymphocytes shared between multiple patients. Because TCRs function as heterodimeric proteins, used an emulsion-based RT-PCR assay link and amplify TCR pairs. Using this on engineered hybridomas,...
Both invasion-promoting MT1-MMP and its physiological inhibitor TIMP-2 play a significant role in tumorigenesis are identified the most aggressive cancers. Despite antiproteolytic effects vitro, clinical data suggest that expression is positively associated with tumor recurrence, thus emphasizing wide-ranging of malignancies. To shed light on this TIMP-2, we report low concentrations by interacting (a specific membrane receptor TIMP-2), induce MEK/ERK signaling cascade fibrosarcoma HT1080...
Abstract Invasion-promoting MT1-MMP is directly linked to tumorigenesis and metastasis. Our studies led us identify those genes, the expression of which universally in multiple tumor types. Genome-wide profiling MT1-MMP–overexpressing versus MT1-MMP–silenced cancer cells a further data mining analysis preexisting database 190 human tumors 14 types 11 correlated firmly with that (P &lt; 0.00001). These genes included regulators energy metabolism (NNT), trafficking membrane fusion (SLCO2A1...
Abstract The integrated stress response (ISR) is an adaptive cellular program that facilitates survival and proliferation under conditions of can be appropriated to contribute the tumorigenesis multiple cancers. ISR activated in various cues tumor microenvironment cell-intrinsic factors but ultimately regulated by four core serine/threonine kinases: GCN2 (activated amino acid deprivation), HRI (in heme deficiency), PERK (unfolded protein stress), PKR (dsRNA infection oxidative stress). These...
Abstract Discovering drug-like molecules for hard-to-target proteins remains a significant challenge. Methyltransferases (DNA, RNA, and protein) are among such targets. At Zafrens, we have developed an ultra-high-throughput discovery platform that leverages large arrays of isolated nano-wells single-bead-single-compound DNA-encoded library (DELs) to uncover target-specific inhibitory impacts on cells. Using combination next-generation sequencing spatially indexed optical imaging biomarkers...
Membrane type-1 matrix metalloproteinase (MT1-MMP) degrades the extracellular matrix, initiates activation pathway of soluble MMPs and regulates functionality cell adhesion signaling receptors, thus playing an important role in many functions. Intracellular transport mechanisms, currently incompletely understood, regulate presentation MT1-MMP at surface. We have focused our efforts on identifying these mechanisms. To understand across cell, we used substitution deletion mutants, trafficking...
Estrogens have many cellular functions, including their interactions with estrogen receptors alpha and beta (ERalpha ERbeta). Earlier, we determined that the estrogen-ER complex stimulates transcriptional activity of matrix metalloproteinase 26 (MMP-26) gene promoter. We then ERbeta is susceptible to MMP-26 proteolysis whereas ERalpha resistant protease. targets NH(2)-terminal region coding for divergent A/B domain responsible ligand-independent transactivation function. As a result,...
MT1-MMP is a key enzyme in cancer cell invasion and metastasis. The activity of cellular regulated by furin-like proprotein convertases, TIMPs, shedding, autoproteolysis, dimerization, exocytosis, endocytosis, recycling. Our data demonstrate that, addition to these already known mechanisms, O-glycosylation its hinge region. Insignificant autolytic degradation characteristic for naturally expressed, glycosylated, MT1-MMP. In turn, extensive degradation, which leads the inactivation protease...
The accurate identification and quantitation of RNA isoforms present in the cancer transcriptome is key for analyses ranging from inference impacts somatic variants to pathway analysis biomarker development subtype discovery. ICGC-TCGA DREAM Somatic Mutation Calling (SMC-RNA) challenge was a crowd-sourced effort benchmark methods isoform quantification fusion detection bulk sequencing (RNA-seq) data. It concluded 2018 with comparison 77 entries 65 on 51 synthetic tumors 32 cell lines...
Abstract Proteases exert control over cell behavior and affect many biological processes by making proteolytic modification of regulatory proteins. The purpose this paper is to describe novel, important functions matrix metalloproteinase (MMP)-26. α1-Antitrypsin (AAT) a serpin, the primary function which regulate activity neutrophil/leukocyte elastase. Insufficient antiprotease because AAT deficiency in lungs contributing factor early-onset emphysema. We recently discovered that efficiently...
Understanding the function of invasion-promoting membrane type-1 matrix metalloproteinase (MT1-MMP) is paramount importance for understanding cancer biology. MT1-MMP synthesized in cells as a latent zymogen that requires cleavage its prodomain to exert proteolytic activity. The mature alphav integrin subunit also generated by endoproteolytic precursor (pro-alphav). Cleavage furin considered be principal event activation both and pro-alphav. To elucidate alternative pathway pro-alphav, we...
Abstract Successful cancer therapies aim to induce selective apoptosis in neoplastic cells. The current suboptimal efficiency and selectivity drugs have therapeutic limitations concomitant side effects. Recently, novel based on the use of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) emerged. TRAIL, a key component natural antitumor immune response, selectively kills many cell types. Earlier studies with recombinant however, revealed its shortcomings including short...
To build a catalog of peptides presented by breast cancer cells, we undertook systematic MHC class I immunoprecipitation followed elution I-loaded in cells. We determined the sequence 3196 ligands representing 1921 proteins from panel 20 cell lines. After removing duplicate peptides, i.e., same peptide eluted more than one line, total number unique was 2740. Of eluted, 1750 had been previously identified, and these, sixteen have shown to be immunogenic. Importantly, half these immunogenic...
An understanding of the regulatory mechanisms that control activity membrane type-1 matrix metalloproteinase (MT1-MMP), a key proteinase in tumor cell invasion, is essential for design potent and safe anti-cancer therapies. A unique proteolytic pathway regulates MT1-MMP at cancer surfaces. The abundance enzymes cells makes it difficult to identify autocatalytic events this pathway. To these events, soluble form MT1-MMP, lacking C-terminal transmembrane cytoplasmic domains, was expressed...
Neoplasms have developed numerous strategies to protect themselves against the host immune system. Membrane type-1 matrix metalloproteinase (MT1-MMP) is strongly associated with many cancer types and up-regulated in aggressive, metastatic neoplasms. During past few years, there has been an increasing appreciation of important, albeit incompletely understood, role MT1-MMP cancer. We discovered, using cell-free cell-based assays vitro, that proteolysis specifically targets C3b, essential...
Abstract An elevated expression of membrane type-1 matrix metalloproteinase (MT1-MMP) is closely associated with multiple malignancies. Recently, we discovered that recycled MT1-MMP was trafficked along the tubulin cytoskeleton into centrosomal compartment and cleaved integral protein pericentrin-2. These events correlated induction chromosome instability aneuploidy in nonmalignant Madine-Darby canine kidney cells. Accordingly, hypothesized an oncogene promotes malignant transformation...