A5103000149- Hedgehog Signaling Pathway Studies
- Cellular transport and secretion
- RNA and protein synthesis mechanisms
- Protein Kinase Regulation and GTPase Signaling
- RNA Research and Splicing
- Microtubule and mitosis dynamics
- Genomic variations and chromosomal abnormalities
- Animal Genetics and Reproduction
- Advanced Synthetic Organic Chemistry
- Erythrocyte Function and Pathophysiology
- Receptor Mechanisms and Signaling
- Cholesterol and Lipid Metabolism
- Glycosylation and Glycoproteins Research
- Pancreatic function and diabetes
- Asymmetric Synthesis and Catalysis
- Peroxisome Proliferator-Activated Receptors
- Developmental Biology and Gene Regulation
- Cancer-related Molecular Pathways
- Cancer and Skin Lesions
- Phosphodiesterase function and regulation
- Sexual Differentiation and Disorders
- Streptococcal Infections and Treatments
- Nonmelanoma Skin Cancer Studies
- Hippo pathway signaling and YAP/TAZ
- Connective tissue disorders research
University of California, San Francisco
2006-2024
University of California System
2024
San Francisco Foundation
2012-2016
University of Nottingham
2011-2012
The University of Queensland
1995-2006
Texas A&M University
2006
Research Institute for Bioscience and Biotechnology
2003
University of California, Los Angeles
1991
Genentech
1989-1991
Doheny Eye Institute
1991
We have isolated cDNA clones from a human placental library that code for low molecular weight GTP-binding protein originally designated Gp (also called G25K). This identification is based on comparisons with the available peptide sequences purified and use of two highly specific anti-peptide antibodies. The predicted amino acid sequence very similar to those various members ras superfamily proteins, including N-, Ki-, Ha-ras proteins (30-35% identical), rho (approximately 50% rac 70%...
Using minor modifications of procedures developed to purify GTP-binding proteins (G-proteins) from rabbit liver, we have purified the major G-proteins present in human placental membranes.One, referred as Gi, is substrate for pertussis toxin-catalyzed ADP-ribosylation and has an a-subunit 41,000 daltons, @-subunit 36,000 35,000 a y-subunit 10,000 daltons.The other protein, G,, was identified by its ability bind guanine nucleotides specifically with high affinity.This activity resolved G, Gi...
We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and this modification occurs at or near the membrane-binding domain. In present study, we identified small G as G25K (formerly termed Gp). demonstrated proteolytic digests of 3H-methylated contained radiolabeled material coeluted with synthetic S-(geranylgeranyl)cysteine methyl ester on reversed-phase HPLC. Further treatment by performic acid oxidation...
The regulation of hedgehog signaling by vesicular trafficking was exemplified the finding that Rab23, a Rab‐GTPase transport protein, is mutated in open brain mice. In this study, localization Rab23 analyzed light and immunoelectron microscopy after expression wild‐type (Rab23‐GFP), constitutively active (Rab23Q68L‐GFP), inactive (Rab23S23N‐GFP) range mammalian cell types. Rab23‐GFP Rab23Q68L‐GFP were predominantly localized to plasma membrane but also associated with intracellular...
Abstract Identification of the GTP-binding proteins from human platelet particulate fractions was attained by their purification via successive column chromatography steps followed amino acid sequencing. To enhance likelihood identifying proteins, two assays were employed to monitor activities: (i) guanosine 5'-(3-O-[35S]thio)triphosphate (GTP gamma S)-binding rapid filtration and ii) [alpha-32P]GTP-binding following sodium dodecyl sulfate-polyacrylamide gel electrophoresis electroblotting...
rap1/Krev-1 is a p21ras-related GTP-binding protein that has been implicated in the reversion of ras-transformed cell phenotype. We have identified GTPase-activating (GAP) specific for rap plasma membranes isolated from differentiated HL60 cells. The GAP activity remained quantitatively associated with membrane following washes buffered 1 M LiCl containing 20 mM EDTA but was solubilized detergents Nonidet P-40 and deoxycholate. On basis size-exclusion chromatography, membrane-associated (rap...
The attenuation of cyclic AMP accumulation occurs by different mechanisms in 1321N1 astrocytoma cells and NG108-15 neuroblastoma X glioma cells. In cells, cholinergic agonists reduce through a Ca2+-dependent activation phosphodiesterase; muscarinic receptor-mediated effects on metabolism occur inhibition adenylate cyclase. goal the current study was to determine whether pharmacological specificities were expressed receptor populations these two cell lines. affinity receptors for...
The significance of the 36,000-35,000-dalton doublet proteins referred to as @-subunit Gproteins remains unresolved.An immunological distinction between 36,000 &e)-and 35,000 (B3& dalton has been reported (Roof, D. J., Apple-
The spatial organization of organelles within a cell is dependent on microtubules. Recently, members the Hook family proteins have been proposed to function in linking We report identification completely novel protein family, Hook‐related (HkRP) from which diverged. Bioinformatic analysis HkRP revealed several conserved domains, including unique C‐terminal domain. central region each comprised an extensive coiled‐coil domain, and N‐terminus contains putative microtubule‐binding This domain...
A guanine-nucleotide-binding protein (G-protein) was purified from cholate extracts of bovine brain membranes by sequential DEAE-Sephacel, Ultrogel AcA-34, heptylamine-Sepharose and Sephadex G-150 chromatography. Guanosine 5′-[gamma-[35S]thio]triphosphate (GTP[35S])-binding activity copurified with a 25,000 Da peptide 35,000-36,000 doublet. Neither pertussis toxin nor cholera catalysed the ADP-ribosylation associated GTP[35S]-binding activity. Photoaffinity labelling 8-azido[gamma-32P]GTP...
Clathrin depletion by ribonucleic acid interference (RNAi) impairs mitotic spindle stability and cytokinesis. Depletion of several clathrin-associated proteins affects centrosome integrity, suggesting a further cell cycle function for clathrin. In this paper, we report that RNAi CHC17 (clathrin heavy chain 17) clathrin, but not the CHC22 clathrin isoform, induced amplification multipolar spindles. To stage within cycle, line expressing SNAP-tagged light chains was generated. Acute...
The clathrin light chain (CLC) subunits participate in several membrane traffic pathways involving both and actin, through binding the actin-organizing huntingtin-interacting proteins (Hip). However, CLCs are dispensable for clathrin-mediated endocytosis of many cargoes. Here we observe that CLC depletion affects cell migration Hip reduces surface expression β1-integrin by interference with recycling following normal inactive β1-integrin. a modified also inhibit appearance gyrating...
An array of 16 enantiomeric pairs chiral phosphate, phosphonate, and phosphinate esters was used to establish the breadth stereoselective discrimination inherent within bacterial phosphotriesterase 15 mutant enzymes. For each substrate, leaving group 4-hydroxyacetophenone while other two groups attached phosphorus core consisted an asymmetric mixture methyl, methoxy, ethyl, ethoxy, isopropoxy, phenyl, phenoxy, cyclohexyl, cyclohexoxy substituents. wild-type enzyme, relative rates hydrolysis...
The abilities of different GTP-binding proteins to serve as phosphosubstrates for the epidermal growth factor (EGF) receptor/tyrosine kinase have been examined in reconstituted phospholipid vesicle systems. During course these studies we discovered that a low molecular mass, high affinity protein from bovine brain (designated 22-kDa protein) served an excellent phosphosubstrate tyrosine-agarose-purified human placental EGF receptor. EGF-stimulated phosphorylation purified occurs on tyrosine...
Significance Clathrin forms coats on vesicles that control receptor endocytosis, thereby influencing cell signaling. The contribution of the clathrin light chain subunits to this process in mice was addressed by deleting gene encoding predominant form (CLCa) present B lymphocytes. CLCa-null have cells with altered antibody production and aberrant development due defects endocytosis signaling receptors influence these processes. Only a subset were found depend chains for their demonstrating...
It has been proposed that beta gamma dimers of signal-transducing G proteins mediate muscarinic activation atrial K+ channels. We examined this hypothesis by testing the effects from four sources (human erythrocytes, human placenta, bovine brain, and retina) on single channel (K+[acetylcholine (ACh)]) currents in inside-out membrane patches adult guinea pig atria. None dimer preparations stimulated K+[ACh] currents; contrary, each inhibited whether were activated with GTP alone...
Loss of heterozygosity occurs frequently on the short arm chromosome 8 in many neoplasms, including colorectal and ovarian cancer. Monochromosome transfer experiments into tumour cell lines have provided functional evidence for a suppressor gene located at 8p22–23. One genes from this region that is expressed by our suppressed hybrids candidate gene, DLC1 (deleted liver cancer), which has homology to rat RhoGAP. We delineated structure used single-stranded conformation polymorphism analysis...
Inactivating mutations in the human patched (PTCH) gene have been identified both familial and sporadic basal cell carcinomas (BCCs). In some tumors detected alleles thereby supporting role of PTCH as a tumor suppressor gene. We analyzed 22/23 coding exons for 44 BCCs, 10 novel nine tumors. two mutant remaining allele was inactivated by loss heterozygosity. Five polymorphisms were also identified. Most variations found C>T substitutions at dipyrimidine sites, previous studies which indicate...