Fluorescence visualization of receptor-bound low density lipoprotein in human fibroblasts.
AuthorsAnderson RG, Goldstein JL, Brown MS
JournalJ Recept Res
PubMed ID6271950
Reconstituted low density lipoprotein: a vehicle for the delivery of hydrophobic fluorescent probes to cells.
AuthorsKrieger M, Smith LC, Anderson RG, Goldstein JL, Kao YJ, Pownall HJ, Gotto AM, Brown MS
JournalJ Supramol Struct
PubMed ID229357
'Previous studies have shown that the cholesteryl ester core of plasma low density lipoprotein (LDL) can be extracted with heptane and replaced with a variety of hydrophobic molecules. In the present report we use this reconstitution technique to incorporate two fluorescent probes, 3-pyrenemethyl-23, 24-dinor-5-cholen-22-oate-3 beta-yl oleate (PMCA oleate) and dioleyl ... More
Isolation of cholesterol-requiring mutant Chinese hamster ovary cells with defects in cleavage of sterol regulatory element-binding proteins at site 1.
AuthorsRawson RB, Cheng D, Brown MS, Goldstein JL
JournalJ Biol Chem
PubMed ID9774448
'The synthesis and uptake of cholesterol requires transcription factors designated sterol regulatory element-binding proteins (SREBPs). SREBPs are bound to membranes in a hairpin orientation with their transcriptionally active NH2-terminal segments facing the cytosol. The NH2-terminal segments are released from membranes by two-step proteolysis initiated by site 1 protease (S1P), which ... More
Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin-coated pit formation.
AuthorsHeuser JE, Anderson RG
JournalJ Cell Biol
PubMed ID2563728
'Two seemingly unrelated experimental treatments inhibit receptor mediated endocytosis: (a) depletion of intracellular K+ (Larkin, J. M., M. S. Brown, J. L. Goldstein, and R. G. W. Anderson. 1983. Cell. 33:273-285); and (b) treatment with hypertonic media (Daukas, G., and S. H. Zigmond. 1985. J. Cell Biol. 101:1673-1679). Since the ... More
Effects of cholesterol in chylomicron remnant models of lipid emulsions on apoE-mediated uptake and cytotoxicity of macrophages.
AuthorsSakurai A, Morita SY, Wakita K, Deharu Y, Nakano M, Handa T
JournalJ Lipid Res
PubMed ID15930510
'Chylomicron remnants have been suggested to be involved in the development of atherosclerosis. To investigate the mechanisms of chylomicron remnant-induced atherosclerosis, we prepared cholesterol (Chol)-containing emulsion particles as models for chylomicron remnants. Chol markedly increased the apolipoprotein E (apoE) binding maximum of emulsions without changing the binding affinity and thereby ... More
Receptor-mediated delivery of photoprotective agents by low-density lipoprotein.
AuthorsMosley ST, Yang YL, Falck JR, Anderson RG
JournalExp Cell Res
PubMed ID6499941
'Low density lipoprotein (LDL) has been used to deliver toxic molecules to cells by receptor-mediated endocytosis. In these studies, the cholesteryl ester core of LDL was replaced with a lipophilic, toxic molecule. We now report that photoprotective azo dyes can be stably incorporated into LDL, and that this reconstituted LDL ... More
Synaptotagmin regulation of coated pit assembly.
Authorsvon Poser C, Zhang JZ, Mineo C, Ding W, Ying Y, Sudhof TC, Anderson RG
JournalJ Biol Chem
PubMed ID10906143
Synaptotagmins bind clathrin AP-2 with high affinity via their second C(2) domain, which indicates they are involved in coated pit function. We now report that expression of synaptotagmins lacking either the second C(2) domain or the entire cytoplasmic region potently inhibit endocytosis. Inhibition was dependent on two intramembrane cysteine residues ... More
Failure to cleave sterol regulatory element-binding proteins (SREBPs) causes cholesterol auxotrophy in Chinese hamster ovary cells with genetic absence of SREBP cleavage-activating protein.
AuthorsRawson RB, DeBose-Boyd R, Goldstein JL, Brown MS
JournalJ Biol Chem
PubMed ID10497220
We describe a line of mutant Chinese hamster ovary cells, designated SRD-13A, that cannot cleave sterol regulatory element-binding proteins (SREBPs) at site 1, due to mutations in the gene encoding SREBP cleavage-activating protein (SCAP). The SRD-13A cells were obtained by two rounds of gamma-irradiation followed first by selection for a ... More
Targeted killing of cultured cells by receptor-dependent photosensitization.
AuthorsMosley ST, Goldstein JL, Brown MS, Falck JR, Anderson RG
JournalProc Natl Acad Sci U S A
PubMed ID6272315
This paper describes a method, designated "receptor-dependent photosensitization," by which the receptor-mediated endocytosis of low density lipoprotein (LDL) can be used to deliver a photosensitizing agent, pyrene, to cultured human and animal cells. The hydrophobic core of LDL is extracted and replaced with pyrene covalently coupled to cholesteryl oleate. This ... More
Calmodulin-dependent enzymes undergo a protein-induced conformational change that is associated with their interactions with calmodulin.
AuthorsHuang S, Carlson GM, Cheung WY
JournalJ Biol Chem
PubMed ID8125988
The anionic hydrophobic (amphipathic) fluorescent probe 2-(p-toluidinyl)-naphthalene-6-sulfonate was used to investigate the surface hydrophobic properties of calmodulin (CaM)-dependent enzymes as follows: calcineurin, myosin light chain kinase, cyclic nucleotide phosphodiesterase, CaM-dependent protein kinase II, and the gamma-subunit of phosphorylase kinase. We found that certain domains of these enzymes that interacted with ... More
Expression cloning of SR-BI, a CD36-related class B scavenger receptor.
AuthorsActon SL, Scherer PE, Lodish HF, Krieger M
JournalJ Biol Chem
PubMed ID7520436
Scavenger receptors are integral membrane proteins that mediate the endocytosis of modified lipoproteins. The first of these to be purified and cloned were the type I and II macrophage scavenger receptors (SR-AI and SR-AII; class A scavenger receptors). Subsequently, the cell surface protein CD36 was shown to bind oxidized low ... More