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S Magnusson

Publications and source records attributed to S Magnusson.

At least 37 records · Page 2Linked to original sources
PubMed1993

Characterization of two distinct pathways of endocytosis of ricin by rat liver endothelial cells.

We have studied the characteristics of internalization and intracellular transport of ricin via two distinct pathways in rat liver endothelial cells (EC), i.e., via binding to mannose receptors and surface galactosyl-residues, respectively. Treatments that inhibit endocytosis from coated pits, i.e., hyperosmolarity and acidification of the cytoplasm, decreased uptake via mannose receptors much more than uptake via galactosyl-residues, indicating that mannose receptors are largely internalized from coated pits, whereas internalization via galactosyl-residues is to a significant extent independent of coated pits. Uptake of ricin via mannose receptors was strongly inhibited by NH4Cl and monensin, and accordingly, NH4Cl protected the cells against ricin intoxication via mannose receptors. On the other hand, uptake via galactosyl-residues was not significantly inhibited by NH4Cl or monensin, and NH4Cl even sensitized the cells to intoxication via this pathway. Brefeldin A, which did not affect ricin uptake, protected the cells against ricin intoxication via either pathway. Protein synthesis in the EC was efficiently inhibited by ricin, even after very short periods of uptake at low ricin concentrations. The onset of protein synthesis inhibition was more rapid upon internalization of ricin via mannose receptors than via galactosyl-residues. Also, ricin internalized via mannose receptors was more efficiently transported from endosomes to lysosomes than ricin internalized via galactosyl-residues. Partial blocking of the galactosyl-binding sites of ricin caused a reduction in the extent of recycling of ricin from endosomes to the cell surface (retroendocytosis), indicating that binding of ricin to membrane galactosyl-residues, which is relatively stable at the slightly acidic pH of endosomes, is an important determinant of the intracellular handling of ricin. We suggest that the observed difference in the transport from endosomes to lysosomes between the two internalization pathways is related to the different stability of the two binding mechanisms at endosomal pH.

Ammonium Chloride↗
PubMed1993

Corticosteroid effect of CPH-82.

A patient suffering from seronegative polyarthritis was treated with CPH-82 (Reumacon). During the treatment he developed a cushingoid appearance and endocrinological abnormalities.

Adrenal Cortex Hormones↗
PubMed1993

Bovine factor XIIa inhibitor.

Bovine factor XIIa inhibitor was purified by an improved method employing affinity for heparin. N-terminal amino acid sequencing revealed a unique sequence without homology to any other known protein sequences. Peptide sequencing, however, showed that a part of the bovine factor XIIa inhibitor was homologous to human C1-inhibitor with a fraction of identical amino acid residues around 70%. Deglycosylation studies and carbohydrate analysis showed the presence of N- and O-linked carbohydrate. Bovine factor XIIa inhibitor did not inhibit plasma kallikrein and trypsin. The reactive site comprised an Arg-Asn bond, and represents the first example of asparagine as a P1' residue in Serpins with well documented inhibitory activity.

Amino Acid Sequence↗
PubMed1993

Receptor-mediated endocytosis of ricin in rat liver endothelial cells. An immunocytochemical study.

The endocytic pathway of ricin in sinusoidal liver endothelial cells (EC) was traced by means of immunocytochemical labeling of ultrathin cryosections. Ricin, a highly mannosylated glycoprotein, is internalized mainly by receptor-mediated endocytosis via the mannose receptor in the EC. Labeling of specimens fixed at different time points after injection of ligand showed that several subcellular compartments are involved in processing of endocytosed ricin. One minute after injection ricin is seen in coated pits, coated vesicles and cisternal-shaped endosomes. After 6 min, the ligand associates with electron-dense, spherical vesicles and electron-lucent vesicles, presumably representing late endosomes. In the same time period we observed labeling in the vicinity of the Golgi stack. At later time points, ricin is increasingly localized in lysosomes. Both late endosomes and lysosomes showed labeling for Igp120, the lysosomal membrane glycoprotein. To compare uptake of ricin with another mannosylated ligand, we coinjected ricin and mannosylated colloidal gold particles (Man-Aun). Man-Au20, injected 24 h before fixation as a marker for late endocytic compartments, was found in two distinct compartments, presumably representing late endosomes and lysosomes. The distribution of ricin and Man-Au10, the latter injected 15 min before fixation, in early endosomes was strikingly different, indicating that the structure of this compartment is important in the process of sorting of ligand and receptor.

Animals↗
PubMed1992

Characterization of retroendocytosis in rat liver parenchymal cells and sinusoidal endothelial cells.

After receptor-mediated endocytosis, internalized ligands may be recycled to the cell surface instead of being routed to lysosomes for degradation, a process termed retroendocytosis. We have investigated the kinetics and extent of retroendocytosis of neoglycoproteins after internalization via two carbohydrate-specific receptors in rat liver cells: galactose receptors in parenchymal cells (PC) and mannose receptors in sinusoidal endothelial cells (EC). Retroendocytosis in both cell types occurred with first-order kinetics, and the rate of recycling of internalized ligands was about 4 times higher in EC than in PC. As the length of the internalization pulse was increased, the extent of subsequent retroendocytosis decreased, indicating that retroendocytosis takes place from a relatively early stage in the endocytic pathway. Furthermore, as the degree of carbohydrate substitution of the neoglycoprotein ligands increased, the affinities of the receptors for the ligands and the extent of ligand retroendocytosis increased. In the EC, the relationship between degree of substitution and extent of retroendocytosis was not immediately apparent, as some of the neoglycoprotein ligands used may also bind to and be internalized by scavenger receptors on the EC, causing a decreased apparent retroendocytosis. However, when this interaction was inhibited, this relationship was restored. We conclude that retroendocytosis mainly occurs because of incomplete dissociation of ligands from receptors before receptor recycling to the cell surface and that the affinities of a receptor for its ligand at the cell surface and in the endosomal environment are major factors in determining the extent of retroendocytosis.

Animals↗
PubMed1992

Complete primary structure of bovine beta 2-glycoprotein I: localization of the disulfide bridges.

The complete primary structure of bovine beta 2-glycoprotein I was determined by a combination of cDNA and peptide sequencing. Bovine beta 2-glycoprotein I was purified from citrated plasma, and by sequencing selected peptides, the complete disulfide bridge patterns of the 11 disulfide bridges were established as well as the positions of the five asparagine-linked carbohydrate groups. Bovine beta 2-glycoprotein I comprises five mutually homologous domains or Short Consensus Repeats, each containing two disulfide bridges, except for the fifth most C-terminal domain which diverges from the Short Consensus Repeat consensus by containing an additional disulfide bridge. In the four N-terminal domains, the first and third and the second and fourth half-cystines are disulfide-linked, while in the fifth domain the first and fourth, the second and fifth, and the third and sixth half-cystines are disulfide-linked.

Amino Acid Sequence↗
PubMed1991

Interactions of ricin with sinusoidal endothelial rat liver cells. Different involvement of two distinct carbohydrate-specific mechanisms in surface binding and internalization.

We have investigated the interactions of the plant toxin ricin with sinusoidal endothelial rat liver cells (EC). In these cells, ricin can be bound and internalized via either cell surface galactosyl residues or mannose receptors. Binding and uptake via galactosyl residues and mannose receptors was studied in the presence of mannan (1 mg/ml) and lactose (50 mM) respectively. Whereas most of the ricin binding was accounted for by cell surface galactosyl residues, uptake of ricin via mannose receptors was much more efficient than uptake via galactosyl residues. Internalized ricin is subject to extensive retroendocytosis (recycling to the cell surface from an early endocytic compartment). Retroendocytosis occurs after internalization of ricin via either pathway and to a much greater extent than for other glycoproteins taken up via mannose receptors of the EC. Hyperosmolarity (150 mM-sucrose), which is known to inhibit endocytosis from coated pits, strongly inhibited ricin uptake via mannose receptors, but had less effect on uptake via galactosyl residues. This suggests that only part of the galactose-specific uptake takes place from coated pits. Protein synthesis in EC was very sensitive to ricin [concn. causing half-maximal inhibition (IC50) = 1.3 x 10(-13) M]. Mannan was slightly more effective than lactose in protecting the EC protein synthesis from ricin toxicity.

Animals↗
PubMed1991

Prevalence of iron deficiency and iron overload in the adult Icelandic population.

The aim of this cross-sectional study was to estimate the prevalence of iron deficiency and overload in the adult population in Iceland, a developed Scandinavian country. The study population consisted of 4240 individuals aged 25-74 years randomly selected from the national roster. Basic hematological, S-iron, S-total iron binding capacity (TIBC), and S-ferritin measurements were obtained on 2588 individuals (61.0%). The results indicated unusually large iron stores in the adult Icelandic population and significantly larger iron stores in the rural compared to the urban population. Iron deficiency was rare except in urban premenopausal women, where 1 in 4 showed evidence of iron deficiency and 3.2% had iron deficiency anemia. Seven patients with hereditary hemochromatosis were identified from a subgroup of 1887 subjects, resulting in a prevalence of 0.37%. Two of the hereditary hemochromatosis patients had been gastrectomized. Measures to improve the iron balance in urban premenopausal women cannot therefore include increased iron fortification of food but must be more directed towards the target group.

Adult↗
PubMed1990

Histidine-rich glycoprotein inhibits contact activation of blood coagulation.

Histidine-rich glycoprotein has been purified from bovine plasma employing two different purification procedures. The first procedure was one-step ion-exchange chromatography using phosphocellulose, while the second procedure involved fractionation using polyethyleneglycol 6000 followed by column chromatography employing CM-Sepharose and heparin-Sepharose. The effect of purified bovine histidine-rich glycoprotein on the contact activation of blood coagulation was studied in human plasma by using as activating surface either an ellagic acid-phospholipid suspension (Cephotest) or sulfatide. Contact activation was monitored by the generation of amidolytic activity towards a synthetic chromogenic substrate (S-2302) for factor XIIa and plasma kallikrein. Bovine histidine-rich glycoprotein inhibits the contact activation induced by both of these activating surfaces.

Amino Acid Sequence↗
PubMed1990

Receptor-mediated endocytosis of ovalbumin by two carbohydrate-specific receptors in rat liver cells. The intracellular transport of ovalbumin to lysosomes is faster in liver endothelial cells than in parenchymal cells.

1. The uptake of ovalbumin (OVA) in rat liver parenchymal cells (PC) and non-parenchymal cells was studied in vivo and in vitro in order to compare the cellular expression of glycoprotein receptors and the kinetics of intracellular transport of ligand endocytosed by these receptors. 2. Ovalbumin was labelled with 125I or with 125I-tyramine-cellobiose (125I-TC). By using 125I-TC-OVA the labelled degradation products were trapped in the cells. 3. 125I-TC-OVA was rapidly cleared from blood mainly by receptor-mediated uptake in the liver. At 30 min after injection, 50% of the ligand was recovered in the liver. The endothelial cells (EC) and the PC were the predominant cell types responsible for uptake. 4. The uptake in PC was strongly inhibited by asialo-orosomucoid (AOM), but not by mannan, indicating that the uptake in these cells was mediated by the galactose receptor and not by the mannose receptor. This finding is compatible with the observation that a proportion of the OVA contains terminal galactose residues in the carbohydrate moiety. 5. In vitro uptake of OVA in cultured EC was saturable and inhibited by mannan, mannose, fructose, N-acetylglucosamine, EDTA or monensin, but not by galactose or AOM. The uptake of OVA in these cells was therefore mediated by the mannose receptor. 6. To label the organelles involved in endocytosis in PC and EC, 125I-TC-OVA was injected intravenously together with an excess of either AOM or mannan. In this way the labelled ligand could be directed selectively to EC or PC respectively. Subcellular fractionation of total liver in sucrose and Nycodenz gradients revealed that in EC the intracellular transport of OVA is so fast that endocytosed ligand accumulates and thus increases the density of the lysosomes. Conversely, in PC transfer of ligand is slower, with the result that accumulation of undegraded ligand in the lysosomes does not occur. These findings are interpreted to mean that in EC the rate-limiting step of handling of endocytosed ligand is intralysosomal degradation, whereas in PC the rate-limiting step is transport of ligand to the lysosomes. 7. Altogether, these findings suggest that endocytosis of OVA by the liver EC and PC is mediated by mannose and galactose receptors respectively, and that the kinetics of intracellular transport of OVA differ in the two cell types.

Animals↗
PubMed1989

Extremely rapid endocytosis mediated by the mannose receptor of sinusoidal endothelial rat liver cells.

Isolated sinusoidal endothelial rat liver cells (EC) in suspension bound and internalized ovalbumin, a mannose-terminated glycoprotein, in a saturable manner. The binding and uptake were Ca2+-dependent and were effectively inhibited by alpha-methyl mannoside and yeast mannan, but not by galactose or asialoglycoproteins. This corresponds to the binding specificity described for the mannose receptor of macrophages and non-parenchymal liver cells. Binding studies indicated a surface pool of 20,000-25,000 mannose receptors per cell, with a dissociation constant of 6 x 10(-8) M. Uptake and degradation of ovalbumin by isolated EC were inhibited by weak bases and ionophores which inhibit acidification of endocytic vesicles and dissociation of receptor-ligand complexes. Cycloheximide had no effect on uptake or degradation. Degradation, but not uptake, was inhibited by leupeptin. We conclude that ovalbumin dissociates from the mannose receptors in the endosomal compartment and the receptors are recycled to the cell surface, while the ovalbumin is directed to the lysosomes for degradation. A fraction of the internalized ovalbumin was recycled intact to the cell surface and escaped degradation (retroendocytosis). The rate of internalization of ovalbumin by isolated EC was very fast, with a Ke (endocytotic rate constant) of 4.12 min-1, which corresponds to a half-life of 10 s for the surface pool of receptor-ligand complexes. To our knowledge, this is the highest Ke reported for a receptor-mediated endocytosis system.

Animals↗
PubMed1988

Contact activation of blood coagulation is inhibited by plasma factor XIII b-chain.

The effect of the purified bovine plasma factor XIII b-chain on contact activation of blood coagulation was studied in human and bovine plasma using either an ellagic acid-phospholipid suspension (Cephotest) or dextran sulfate as activating surface. Contact activation was monitored by the generation of amidolytic activity towards a synthetic chromogenic substrate (S-2302) for factor XIIa and plasma kallikrein. The factor XIII b-chain, which is released from tetrameric factor XIII (a2b2) in the late stages of blood coagulation, inhibits contact activation induced by both activation surfaces mentioned. It was shown that a 5 min preincubation of the factor XIII b-chain with the activation surface increases its inhibitory effect. Light scattering measurements indicated a concurrent binding of the factor XIII b-chain to the Cephotest material. Because factor XIII (a2b2) itself had no such inhibitory activity, the present finding that the factor XIII b-chain inhibits contact activation may point to a novel feed-back inhibition mechanism of blood coagulation.

Animals↗
PubMed1987

Primary structure of tetranectin, a plasminogen kringle 4 binding plasma protein: homology with asialoglycoprotein receptors and cartilage proteoglycan core protein.

Tetranectin binds to plasminogen and to isolated kringle 4 [Clemmensen, I., Petersen, L. C., & Kluft, C. (1986) Eur. J. Biochem. 156, 327-333], apparently to its lysine-binding site. Each of the four identical chains consists of 181 amino acid residues. The three intrachain disulfide bonds connect Cys residues 50-60, 77-176, and 152-168. The tetranectin sequence is homologous (17-24% identical positions) with those parts of the asialoglycoprotein receptor family that are considered to be extracellular. Tetranectin has no structures corresponding to those parts of the receptors considered to be intracellular and membrane anchoring. The sequence of tetranectin is also homologous (22-23% identical positions) with the C-terminal globular domain of the core protein of the cartilage proteoglycan. All six Cys residues in tetranectin are located at positions that are also Cys residues in this proteoglycan. Therefore, a plausible disulfide bond pattern can now be proposed for both the asialoglycoprotein receptors and the C-terminal domain of the proteoglycan core protein. No covalently bound carbohydrate has been found.

Aggrecans↗
PubMed1987

Disulphide bridges of bovine factor X.

Evidence is presented for the disulphide bridges in bovine Factor X. The protein was degraded by chemical and enzymic means, and all 12 disulphide bridges were isolated in separate peptides except for bridges nos. 6/7 in the light chain. All the disulphide bridges were found to be in positions corresponding to those found in other homologous domains. This report is the first verification of an epidermal-growth-factor-homologous domain having the same disulphide-bonding pattern as that found in mouse epidermal growth factor.

Amino Acids↗
PubMed1986

Complete primary structure of bovine plasma fibronectin.

The primary structure of the 2265 residues of bovine plasma fibronectin has been completed. The new sequences reported in this paper are residues 600-868 (269 residues), 1138-1217 (80 residues), 1518-1599 (82 residues) and 1868-2061 (194 residues). These sequences constitute six type III homology units and two non-homologous connecting strands. Thus, there are fifteen type III homology units in plasma fibronectin. Evidence for two of the three splice variants found in rat liver cells [Schwarzbauer et al. (1983) Cell 35, 421-431] was obtained. No indication of the 'extra' type III domain present in some human fibroblast fibronectins [Kornblihtt et al. (1984) EMBO J. 3, 221-226] was found. Three carbohydrate groups (two glucosamine-based and one galactosamine-based) were identified, giving a total of eight carbohydrate groups in the longest splice variant of bovine plasma fibronectin. A second free sulfhydryl group (cysteine) was identified. This cysteine, like the first, is in a type III homology unit. HPLC patterns of peptides obtained from the C-terminal 6-kDa fragment suggested that the interchain bridge pattern of fibronectin is antiparallel. The bovine plasma fibronectin sequence is highly homologous to the human cellular fibronectin sequence deduced from the cDNA sequence [Kornblihtt et al. (1985) EMBO J. 4, 1755-1759].

Amino Acid Sequence↗
PubMed1986

Donor and acceptor splice signals within an exon of the human fibronectin gene: a new type of differential splicing.

We have sequenced that area of a human fibronectin gene clone which codes for a connecting strand separating the last two areas of the type III homology. The gene has a complex exon with two 'AG' acceptor sites. One of these can be used (exon subdivision). In addition 93 basepairs inside the exon are sometimes spliced out as an intron. This is the third differential splicing found in the fibronectin gene transcript and it represents a new type of differential splicing.

Base Composition↗
PubMed1986

Purification and complete primary structures of the heparin-, cell-, and DNA-binding domains of bovine plasma fibronectin.

The complete amino acid sequences of the heparin-, cell- and DNA-binding domains of bovine plasma fibronectin have been determined. The fragments were generated from the 170-kDa central plasmic fragment by extensive digestion with chymotrypsin, and they contain 268, 300 and 269 amino acid residues, respectively. No half-cystines or cysteines were found in these sequences. A glucosamine-based oligosaccharide group is attached to Asn-108 in the sequence of the DNA-binding domain. Only one of the three types of internal homology found in fibronectin [Petersen et al. (1983) Proc. Natl Acad. Sci. USA 80, 137-141], namely the type III homology, occurs in these three fragments, and each of them consists of approximately three stretches of this type III homology. Part of the arrangement of peptides was derived by comparison with the partial cDNA sequence for human fibronectin recently reported [Kornblihtt et al. (1984) Nucleic Acids Res. 12, 5853-5868].

Amino Acid Sequence↗