A5030103232- Cellular transport and secretion
- Endoplasmic Reticulum Stress and Disease
- Cell death mechanisms and regulation
- Autophagy in Disease and Therapy
- HIV Research and Treatment
- Lysosomal Storage Disorders Research
- Mitochondrial Function and Pathology
- Genetics and Neurodevelopmental Disorders
- Peptidase Inhibition and Analysis
- Protease and Inhibitor Mechanisms
- Immune Cell Function and Interaction
- Sirtuins and Resveratrol in Medicine
- Ubiquitin and proteasome pathways
- Herpesvirus Infections and Treatments
- Cytomegalovirus and herpesvirus research
- Signaling Pathways in Disease
- Genomics and Rare Diseases
- RNA modifications and cancer
- HIV/AIDS drug development and treatment
- Retinal Development and Disorders
- Adipose Tissue and Metabolism
- Cell Adhesion Molecules Research
- Complement system in diseases
- Enzyme Structure and Function
- Pancreatic function and diabetes
University of Pittsburgh
2015-2024
UPMC Hillman Cancer Center
2017-2024
Pittsburg State University
2020
Oregon Health & Science University
2001-2012
Vollum Institute
2003-2012
Yale University
2006
Vaccine and Gene Therapy Institute
2006
Fox Chase Cancer Center
2001
Shriners Hospitals for Children - Portland
2001
University of California, San Diego
2000
Previous work demonstrated that human furin is a predominantly Golgi membrane-localized endoprotease can efficiently process precursor proteins at paired basic residues (-Lys-Arg- or -Arg-Arg-) in transfected cells. Anion-exchange chromatography of culture supernatant from cells expressing soluble truncated form resulted greatly enriched preparation the (approximately 70% pure as determined by protein staining). Enzymatic studies show calcium-dependent (K0.5 = 200 microM) serine which has...
Proteolytic cleavage of the protective antigen (PA) protein anthrax toxin at residues 164-167 is necessary for toxic activity. Cleavage by a cellular protease this sequence, Arg-Lys-Lys-Arg, normally follows binding PA to cell surface receptor. We attempted identify determining its sequence specificity and catalytic properties. Semi-random cassette mutagenesis was used generate mutants with replacements Arg, Lys, Ser, or Asn. Analysis 19 mutant proteins suggested that lethal factor-dependent...
Extracts from BSC-40 cells infected with vaccinia recombinants expressing either the yeast KEX2 prohormone endoprotease or a human structural homologue (fur gene product) contained an elevated level of membrane-associated endoproteolytic activity that could cleave at pairs basic amino acids (-LysArg- and -ArgArg-). The fur-directed (furin) shared many properties Kex2p including pH 7.3 requirement for calcium. By using antifurin antibodies, immunoblot analysis detected two furin translation...
The important role of furin in the proteolytic activation many pathogenic molecules has made this endoprotease a target for development potent and selective antiproteolytic agents. Here, we demonstrate utility protein-based inhibitor α 1 -antitrypsin Portland (α -PDX) as an antipathogenic agent that can be used prophylactically to block furin-dependent cell killing by Pseudomonas exotoxin A. Biochemical analysis specificity bacterially expressed His- FLAG-tagged -PDX -PDX/hf) revealed...
Calnexin is an endoplasmic reticulum (ER) lectin that mediates protein folding on the rough ER. also interacts with ER calcium pumps localize to mitochondria-associated membrane (MAM). Depending homeostasis, varying amounts of calnexin target plasma membrane. However, no regulated sorting mechanism so far known for calnexin. Our results now describe how interaction cytosolic PACS-2 distributes between ER, MAM, and Under control conditions, more than 80% localizes majority MAM. knockdown...
Pro-protein convertases such as furin are expressed in many human tumor lines and primary tumors. Furin processes stromelysin-3, membrane type 1 matrix metalloproteinase (MMPs) involved cell invasiveness, well growth factors transforming factor β1. Evaluation of expression head neck squamous carcinoma (HNSCC) cells exhibiting different invasive ability showed that overexpression correlated with their respective invasiveness. The use a selective inhibitor, alpha 1-PDX (PDX) was studied three...
Aerolysin is secreted as an inactive dimeric precursor by the bacterium <i>Aeromonas hydrophila</i>. Proteolytic cleavage within a mobile loop near C terminus of protoxin required for oligomerization and channel formation. This contains sequence KVRRAR<sup>432</sup>, which should be recognized mammalian proprotein convertases such furin, PACE4, PC5/6A. Here we show that these three proteases cleave proaerolysin after Arg-432 <i>in vitro</i>, yielding active toxin. We also investigated...
Furin catalyzes the proteolytic maturation of many proproteins within trans-Golgi network (TGN)/endosomal system. Furin's cytosolic domain (cd) directs both compartmentalization to and transit between its manifold processing compartments (i.e., TGN/biosynthetic pathway, cell surface, endosomes). Here we report identification first furin cd sorting protein, ABP-280 (nonmuscle filamin), an actin gelation protein. The was used as bait in a yeast two-hybrid screen identify furin-binding Binding...
The propeptide of furin has multiple roles in guiding the activation endoprotease vivo. 83-residue N-terminal is autoproteolytically excised endoplasmic reticulum (ER) at consensus site, -Arg104-Thr-Lys-Arg107↓-, but remains bound to as a potent autoinhibitor. Furin lacking ER-retained and proteolytically inactive. Co-expression with propeptide, however, restores trans-Golgi network (TGN) localization enzyme activity, indicating that an intramolecular chaperone. Blocking this step results...
Proteolytic maturation of proBMP-4 is required to generate an active signaling molecule. We show that cleaved by furin in a sequential manner. Cleavage at consensus site adjacent the mature ligand domain allows for subsequent cleavage upstream nonconsensus within prodomain. BMP-4 synthesized from precursor which noncleavable less active, signals shorter range, and accumulates lower levels than does native precursor. Conversely, both sites are rapidly more over greater range. Differential use...
ABSTRACT The genomes of several poxviruses contain open reading frames with homology to the K3 and K5 genes Kaposi's sarcoma-associated herpesvirus (KSHV) gene murine gammaherpesvirus 68, which target major histocompatibility complex class I (MHC-I) as well costimulatory molecules for proteasomal or lysosomal degradation. homologous product myxomavirus (MV), M153R, was recently shown reduce cell surface expression MHC-I. In addition, normal MHC-I observed in cells infected MV lacking M153R...
Mitochondria-associated ER membranes (MAMs) are crucial for lipid transport and synthesis, calcium exchange, mitochondrial functions, they also act as signaling platforms. These contact sites play a critical role in the decision between autophagy apoptosis with far reaching implications cell fate. Vascular smooth muscle (VSMC) accelerates atherogenesis progression of advanced lesions, leading to atherosclerotic plaque vulnerability medial degeneration. Though successful damaged mitochondria...
X-linked hypohidrotic ectodermal dysplasia (XLHED) is a heritable disorder of the ED-1 gene disrupting morphogenesis structures. The product, ectodysplasin-A (EDA), tumor necrosis factor (TNF) family member and synthesized as membrane-anchored precursor protein with TNF core motif located in C-terminal domain. stalk region EDA contains sequence -Arg-Val-Arg- Arg 156 -Asn-Lys-Arg 159 -, representing overlapping consensus cleavage sites (Arg-X-Lys/Arg-Arg ↓ ) for proprotein convertase furin....