A5049671948- Cellular transport and secretion
- Glycosylation and Glycoproteins Research
- Lysosomal Storage Disorders Research
- Calcium signaling and nucleotide metabolism
- Advanced Proteomics Techniques and Applications
- Erythrocyte Function and Pathophysiology
- Monoclonal and Polyclonal Antibodies Research
- Advanced biosensing and bioanalysis techniques
- Adenosine and Purinergic Signaling
- Biochemical and Structural Characterization
- Cell death mechanisms and regulation
- Advanced Biosensing Techniques and Applications
- Signaling Pathways in Disease
- Brain Metastases and Treatment
- RNA modifications and cancer
- Cancer, Stress, Anesthesia, and Immune Response
- RNA Interference and Gene Delivery
- Mechanisms of cancer metastasis
- Carbohydrate Chemistry and Synthesis
- Protease and Inhibitor Mechanisms
- Cancer Research and Treatments
- Ubiquitin and proteasome pathways
- Polysaccharides and Plant Cell Walls
- ATP Synthase and ATPases Research
- Pharmacological Effects and Assays
Tufts University
1998-2019
University of North Carolina Health Care
2019
National Institutes of Health
1982-2019
Northwestern University
2019
University of Kansas
2019
Leidos (United States)
2019
Leidos Biomedical Research Inc. (United States)
2019
Pfizer (United States)
2011
Tufts Medical Center
2010
Weatherford College
1995
Abstract Background The 4T1 mouse mammary tumor cell line is one of only a few breast cancer models with the capacity to metastasize efficiently sites affected in human cancer. Here we describe two lines modified facilitate analysis growth and metastasis evaluation gene function vivo . New information regarding involvement innate acquired immunity other characteristics model relevant its use study late stage are reported. Methods were engineered for stable expression firefly luciferase allow...
It has previously been shown that highly invasive MDA-MB231 human breast cancer cells express vacuolar proton-translocating ATPase (V-ATPases) at the cell surface, whereas poorly MCF7 line does not. Bafilomycin, a specific V-ATPase inhibitor, reduces in vitro invasion of MB231 but not cells. Targeting V-ATPases to different cellular membranes is controlled by isoforms subunit a. mRNA levels for were measured and using quantitative reverse transcription-PCR. The results show although all four...
A receptor that binds the phosphomannosyl recognition marker of bovine testicular beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) was isolated from liver membranes. The extracted crude plasma membrane preparations with Triton X-100 and immunoprecipitated as a receptor--beta-galactosidase complex anti-beta-galactosidase. dissociated precipitate mannose 6-phosphate, labeled 125I, purified on beta-galactosidase-Sepharose 4B affinity matrix. quantitative binding assay...
The natural history of the mannose 6-phosphate receptor was examined by radiolabeling cells in monolayers or suspension; isolated immuno- affinity precipitation followed polyacrylamide gel electrophoresis. found to contain asparagine-linked oligosaccharide chains and phosphorylated serine residues. Newly made sensitive endo-beta-N-acetylglucosaminidase H (endo-H) slowly converted a mature endo-H resistant form; phosphate on only. had an apparent molecular weight 215,000 at all times, as...
Bovine testicular β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) is rapidly and selectively assimilated by human skin fibroblasts. The assimilation of the enzyme strongly inhibited mannose 6-phosphate a glycoprotein fraction isolated from bovine testes (glycoprotein inhibitors). These results suggest that inhibitors have common recognition marker contains 6-phosphate. presence phosphate in was demonstrated acid hydrolysis glycoproteins to liberate followed reduction with NaB...
A compound that reduces the prevalence of perinucleolar compartment in cancer cells inhibits metastasis vivo.
Abstract The insulin-like growth factor-II/mannose 6-phosphate receptor which targets acid hydrolases to lysosomes, has two different binding sites, one for the mannose (Man-6-P) recognition marker on lysosomal enzymes and other factor-II (IGF-II). We have asked whether IGF-II can regulate cellular uptake of enzyme 125I-beta-galactosidase by modulating IGF-II/Man-6-P receptor. first isolated high affinity chromatography an receptor-Sepharose column. Specific (mannose 6-phosphate-inhibitable)...
Triple negative breast cancer (TNBC) is a heterogeneous disease that has poor prognosis and limited treatment options. Chemokine receptor interactions are important modulators of metastasis; however, it now recognized quantitative surface expression one chemokine receptor, CXCR4, may not directly correlate with metastasis its functional activity in better inform tumor pathogenicity. G protein coupled kinase 3 (GRK3) regulator CXCR4 activity, we show GRK correlates tumorigenicity, molecular...
Studies in recent years have indicated that secretion of certain lysosomal hydrolases can be enhanced under various conditions. One such protein, the major excreted protein (MEP) Kirsten virus-transformed NIH 3T3 (KNIH) fibroblasts, is a cysteine protease whose synthesis and are affected by viral transformation growth factors. We been studying transport MEP order to understand better mechanisms responsible for regulation enzyme secretion. Synthesis KNIH cells was found 25-fold greater than...
Abstract The cation-independent mannose 6-phosphate receptor (MPRCI) functions in the packaging of both newly made and extracellular lysosomal enzymes into lysosomes. subcellular location MPRCI reflects these two functions; is found Golgi complex, endosomes, on cell surface. To learn about intracellular pathway followed by surface to study relationship between pools, we examined entry compartments that contain sialyltransferase. Sialic acid was removed from surface-labeled K562 cultured...
This report summarizes studies concerning the role of lysosomal protein: Man‐6‐P receptor and describes some recent data on its biosynthesis cellular translocation. The functions both in Golgi apparatus (or GERL) cell surface where it binds proteins mediates their transport to lysosomes. Consistent with dual role, several types has been localized plasma membrane cisternae, clathrin‐coated structures at locations, vesicles characteristic endosomes or CURL. Biosynthetic have shown that...
We have found that the major excreted protein (MEP) of transformed mouse fibroblasts, a phosphoglycoprotein Mr = 35,000, carries mannose 6-phosphate
Using ultrastructural immunocytochemistry and antibodies directed against bovine liver phosphomannosyl (PM) receptor, we have localized the receptor in Chinese hamster ovary (CHO) cells. The majority of was found within cell. Only a small fraction on surface most it clustered coated pits. Because these cells contain endogenous ligands for not possible to determine if this state dependent occupancy receptor. bulk cell's endoplasmic reticulum, nuclear envelope, Golgi system. Most localization...
The major excreted protein (MEP) of transformed mouse fibroblasts is the lysosomal protease, cathepsin L. MEP also secreted by untransformed cells in response to growth factors and tumor promoters, thought play a role cell transformation. To determine relationship between synthesis secretion, we have examined these events PDGF-treated NIH 3T3 cells. PDGF enhanced caused diversion from delivery pathway secretory pathway. These two effects were found be regulated independently at various times...
Previous studies have shown that lysine residues on the surface of cathepsins and other lysosomal proteins are a shared component recognition structure involved in mannose phosphorylation. In this study, involvement specific phosphorylation cathepsin D was explored by site-directed mutagenesis. Mutation two mature portion protein, Lys-203 Lys-293, cooperated to inhibit 70%. Other positively charged could not substitute for at these positions, comparison thermal denaturation curves wild type...
Abstract We developed surface proteome signatures (SPS) for identification of new biomarkers playing a role in cancer drug resistance. SPS compares antigen expression different cell lines by immunocytochemistry phage display antibody library directed to antigens HT1080 fibrosarcoma cells. applied compare the proteomes two epithelial derived lines, MCF7 and NCI/ADR-RES, which is resistant because overexpression P-glycoprotein (P-gp) efflux pump. Surface proteomic profiling identified CD44 as...
The recognition of lysosomal enzymes by UDP-GlcNAc:lysosomal-enzyme GlcNAc-1-phosphotransferase (phosphotransferase) is mediated a protein structure on enzymes. It has been previously demonstrated that lysine residues are required for phosphorylation procathepsin L and common feature the site many proteins. In this work, was further defined identification region containing critical involved. Removal cathepsin propeptide low pH-induced autocatalytic processing abolished phosphorylation....
In cultured mouse fibroblasts, secretion of the lysosomal cysteine protease, MEP (major excreted protein) is regulated by growth factors and viral transformation. The ability this protein to be has been attributed its intrinsic low affinity for cation-independent mannose 6-phosphate (Man-6-P) receptor (Dong, J., Prence, E. M., Sahagian, G. (1989) J. Biol. Chem. 264, 7377-7383). study, basis was examined. Chromatography on a Man-6-P matrix used assess relative affinities Man-6-P-containing...
The mannose 6-phosphate (Man-6-P) receptor is an integral membrane glycoprotein which mediates intracellular transport and receptor-mediated endocytosis of lysosomal proteins. Clathrin-coated vesicles, have been shown to be significantly involved in these processes, also a major subcellular site the receptor. In order define orientation Man-6-P within coated vesicle membrane, highly purified preparations vesicles were prepared from bovine brain employing D2O/sucrose gradient centrifugation...