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Results for “Extracellular Traps”

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PubMed1994

Rat long-term bone marrow culture as a model for transient production of multinucleated giant cells into suspension culture fraction and for steady mineralization process of adherent stromal cells.

Under in vitro conditions rat bone marrow stromal cells induced into osteogenic differentiation by beta-glycerophosphate, exogenous ascorbic acid and dexamethasone are able to produce a mineralized matrix. Here we describe adult rat long-term bone marrow cultures (LTBMC), deprived of these substances. Mineralization process in stromal adherent cells occurred after 1 month of incubation and was proved by means of X-ray electron microprobe. The high content of sulfur in the extracellular matrix surrounding the mineralized nodules suggests non-distrophic pathway of Ca/P deposition. During the first 2 weeks of incubation the extensive production of multinucleated tartrate-resistant acid phosphatase (TRAP) positive and TRAP negative giant cells occurred into the suspension fraction of the cultures. Rat LTBMC may serve as a model for the investigation of differentiation of osteoblastic progenitors and their interactions with multinucleated TRAP positive cells.

Animals↗

PubMed2005

Comprehensive analysis of protein folding activation thermodynamics reveals a universal behavior violated by kinetically stable proteases.

Alpha-lytic protease (alpha LP) and Streptomyces griseus protease B (SGPB) are two extracellular serine proteases whose folding is absolutely dependent on the existence of their companion pro regions. Moreover, the native states of these proteins are, at best, marginally stable, with the apparent stability resulting from being kinetically trapped in the native state by large barriers to unfolding. Here, in an effort to understand the physical properties that distinguish kinetically and thermodynamically stable proteins, we study the temperature-dependences of the folding and unfolding kinetics of alpha LP and SGPB without their pro regions, and compare their behavior to a comprehensive set of other proteins. For the folding activation thermodynamics, we find some remarkable universal behaviors in the thermodynamically stable proteins that are violated dramatically by alpha LP. Despite significant variations in deltaC(P,F)++, the maximal folding speed occurs within the narrow biological temperature range for all proteins, except for alpha LP, with its maximal folding speed shifted lower by 200 K. This implies evolutionary pressures on folding speed for typical proteins, but not for alpha LP. In addition, the folding free energy barrier in the biological temperature range for most proteins is predominantly enthalpic, but purely entropic for alpha LP. The unfolding of alpha LP and SGPB is distinguished by three properties: a remarkably large deltaC(P,U)++, a very high deltaG(U)++, and a maximum deltaG(u)++ at the optimal growth temperature for the organism. While other proteins display each of these traits to some approximation, the simultaneous optimization of all three occurs only in the kinetically stable proteins, and appears to be required to maximize their unfolding cooperativity, by suppressing local unfolding events, and slowing the rate of global unfolding. Together, these properties extend the lifetime of these enzymes in the highly proteolytic extracellular environment. Attaining such functional properties seems possible only through the gross perturbation of the folding thermodynamics, which in turn has required the co-evolution of pro regions as folding catalysts.

Bacterial Proteins↗

PubMed1975

Glomerular filtration rate and microsphere distributions in single nephron of rat kidney.

In control non diuretic (ND) and in salt-loaded (SL) rats, both the microsphere technique and the 14C ferrocyanide infusion technique were used to determine the distribution of microspheres in single glomeruli and the SNGFR of the corresponding nephrons. A sample of microspheres with a diameter averaging 11.0 +/- 2.6 mu SD was selected from a 15 +/- 5 mu unlabelled batch. In each rat, three million of these microspheres were injected through the left carotid artery. The microspheres were directly counted under microscopic observation in the glomeruli of the nephrons which were microdissected to determine the SNGFR value. The number of microspheres per glomerulus for a given kidney generally varied from 0 to 8 and was independent to the SNGFR value. The diameter of the microspheres trapped was constant in all the animals. The mean number of microspheres for superificial (S) and juxtamedullary (JM) nephrons was, (ND); S = 1.99 +/- 0.48 SE., n = 5; JM = 3.02 +/- 0.51 SE, n = 5; P less than 0.02, (SL): S = 3.75 +/- 0.53 SE, n = 6; JM = 2.86 +/- 0.33 SE, n = 6; P less than 0.05. This distribution was directly related to that of SNGFR in ND rats (S = 39.0 +/- 6.1 SD and JM = 49.5 +/- 10.3 nl/min) but not in SL rats (S = 50.9 +/- 6.1 and JM = 66.9 +/- 10.0 nl/min). In conclusion, the microsphere technique described in the present paper, appears more suitable for investigating single glomerular blood flow since the number and the size of the microspheres trapped are directly determined at the level of the glomerulus.

Animals↗

PubMed1996

The crystal structure of a five-stranded coiled coil in COMP: a prototype ion channel?

Oligomerization by the formation of alpha-helical bundles is common in many proteins. The crystal structure of a parallel pentameric coiled coil, constituting the oligomerization domain in the cartilage oligomeric matrix protein (COMP), was determined at 2.05 angstroms resolution. The same structure probably occurs in two other extracellular matrix proteins, thrombospondins 3 and 4. Complementary hydrophobic interactions and conserved disulfide bridges between the alpha helices result in a thermostable structure with unusual properties. The long hydrophobic axial pore is filled with water molecules but can also accommodate small apolar groups. An "ion trap" is formed inside the pore by a ring of conserved glutamines, which binds chloride and probably other monatomic anions. The oligomerization domain of COMP has marked similarities with proposed models of the pentameric transmembrane ion channels in phospholamban and the acetylcholine receptor.

Amino Acid Sequence↗

PubMed2002

Na(+)/H(+) exchange subtype 1 inhibition during extracellular acidification and hypoxia in glioma cells.

Lactacidosis is a common feature of ischaemic brain tissue, but its role in ischaemic neuropathology is still not fully understood. Na(+)/H(+) exchange, a mechanism involved in the regulation of intracellular pH (pH(i)), is activated by low pH(i). The role of Na(+)/H(+) exchange subtype 1 was investigated during extracellular acidification and subsequent pH recovery in the absence and presence of (4-isopropyl-3-methylsulphonyl-benzoyl)-guanidine methanesulfonate (HOE642, Cariporid), a new selective and powerful inhibitor of the Na(+)/H(+) exchanger subtype 1 (NHE-1). It was compared for normoxia and hypoxia in two glioma cell lines (C6 and F98). pH(i) was monitored by fluorescence spectroscopy using the intracellularly trapped pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Alterations in glial cell metabolism were characterized using high-resolution (1)H, (13)C and (31)P NMR spectroscopy of perchloric acid extracts. NHE-1 contributed to glial pH regulation, especially at pathologically low pH(i) values. NHE-1 inhibition with HOE642 during acidification caused exacerbated metabolic disorders which were prolonged during extracellular pH recovery. However, NHE-1 inhibition during hypoxia protected the energy state of glial cells.

Acids↗

PubMed2006

Ab initio computational modeling of long loops in G-protein coupled receptors.

A newly developed approach for predicting the structure of segments that connect known elements of secondary structure in proteins has been applied to some of the longer loops in the G-protein coupled receptors (GPCRs) rhodopsin and the dopamine receptor D2R. The algorithm uses Monte Carlo (MC) simulation in a temperature annealing protocol combined with a scaled collective variables (SCV) technique to search conformation space for loop structures that could belong to the native ensemble. Except for rhodopsin, structural information is only available for the transmembrane helices (TMHs), and therefore the usual approach of finding a single conformation of lowest energy has to be abandoned. Instead the MC search aims to find the ensemble located at the absolute minimum free energy, i.e., the native ensemble. It is assumed that structures in the native ensemble can be found by an MC search starting from any conformation in the native funnel. The hypothesis is that native structures are trapped in this part of conformational space because of the high-energy barriers that surround the native funnel. In this work it is shown that the crystal structure of the second extracellular loop (e2) of rhodopsin is a member of this loop's native ensemble. In contrast, the crystal structure of the third intracellular loop is quite different in the different crystal structures that have been reported. Our calculations indicate, that of three crystal structures examined, two show features characteristic of native ensembles while the other one does not. Finally the protocol is used to calculate the structure of the e2 loop in D2R. Here, the crystal structure is not known, but it is shown that several side chains that are involved in interaction with a class of substituted benzamides assume conformations that point into the active site. Thus, they are poised to interact with the incoming ligand.

Amino Acid Sequence↗

PubMed1995

Chloride is preferentially accumulated in a subpopulation of dendrites and periglomerular cells of the main olfactory bulb in adult rats.

GABA is predominantly an inhibitory transmitter. Mediated by GABAA receptors, GABA opens chloride channels, induces a passive flux of chloride ions, which is usually directed from extracellular to intracellular space, and hyperpolarizes postsynaptic neurons. Recent electrophysiological data suggested that GABA may also depolarize neurons and exert excitatory actions. However, it remained unclear whether excitatory GABA effects are based on reversed transmembrane chloride gradient due to modifications in extracellular or intracellular chloride concentrations. Here, the first histochemical evidence is provided for local redistribution of chloride in the CNS of healthy adult rats. Olfactory bulbs were examined using freeze substitution, silver trapping of chloride and intensification techniques at light and electron microscopic level. The chloride content of precipitates was evidenced by electron spectroscopic imaging using a CEM 902 (Zeiss) electron microscope. Chloride concentration was high in a subpopulation of some periglomerular cell bodies and isolated dendritic profiles, while it seemed to be very low in certain parts of the glomerular neuropil including intercellular clefts. Data suggest that reversed chloride gradients can be demonstrated by cytochemical methods, and may be responsible for excitatory GABA effects on selected periglomerular neurons and dendrites in the olfactory glomeruli. Conditions leading to chloride redistribution in the CNS of normal adult rats remain to be determined.

Animals↗

PubMed2000

The possible role of gradual accumulation of copper, cadmium, lead and iron and gradual depletion of zinc, magnesium, selenium, vitamins B2, B6, D, and E and essential fatty acids in multiple sclerosis.

Multiple sclerosis (MS) has a much higher incidence among caucasians that in any other race. Furthermore: females are much more susceptible than males and white females living in colder, wetter areas are much more susceptible than those living in warmer areas. On the other hand, menstruating women have increased copper (Cu) absorption and half-life, so they tend to accumulate more Cu than males. Moreover, rapidly growing girls have an increased demand for zinc (Zn), but their rapidly decreasing production of melatonin results in impaired Zn absorption, which is exacerbated by the high Cu levels. The low Zn levels result in deficient CuZnSuperoxide dismutase (CuZnSOD), which in turn leads to increased levels of superoxide. Menstruating females also often present with low magnesium (Mg) and vitamin B6 levels. Vitamin B6 moderates intracellular nitric oxide (NO) production and extracellular Mg is required for NO release from the cell, so that a deficiency of these nutrients results in increased NO production in the cell and reduced release from the cell. The trapped NO combines with superoxide to form peroxinitrite, an extremely powerful free radical that leads to the myelin damage of MS. Iron (Fe), molybdenum (Mo) and cadmium (Cd) accumulation also increase superoxide production. Which explains MS in males, who tend to accumulate Fe much faster and Cu much less rapidly than females. Since vitamin D is paramount for Mg absorption, the much reduced exposure to sunlight in the higher latitudes may account for the higher incidence in these areas. Moreover, vitamin B2 is a cofactor for xanthine oxidase, and its deficiency exacerbates the low levels of uric acid caused by high Cu levels, resulting in myelin degeneration. Finally Selenium (Se) and vitamin E prevent lipid peroxidation and EPA and DHA upregulate CuZnSOD. Therefore, supplementation with 100 mg MG, 25 mg vit B6, 10 mg vit B2, 15 mg Zn and 400 IU vit D and E, 100 microg Se, 180 mg EPA and 120 mg DHA per day between 14 and 16 years of age may prevent MS.

Adolescent↗

PubMed1992

Estimation of Ammonium Ion Distribution between Cytoplasm and Vacuole Using Nuclear Magnetic Resonance Spectroscopy.

Evidence is presented that intracellular ammonium is trapped in vacuoles of maize (Zea mays L.) root tips because of rapid movement of ammonia between cytoplasm and vacuoles. The concentration of cytoplasmic ammonium is estimated to be <15 mum at extracellular ammonium concentrations up to 1 mm. The implications for pathways of ammonium assimilation are discussed.

Journal Article↗

PubMed1988

pH regulatory mechanisms in rat pancreatic ductal cells.

The mechanisms underlying regulation of intracellular pH (pHi) by rat pancreatic duct cells were studied by use of the pH-sensitive, fluorescent, cytoplasmically trapped dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Cells exhibited a mean pHi of 7.18 +/- 0.14 in bicarbonate-buffered medium, as calculated from the BCECF fluorescence ratio. Removal of extracellular Na (Nao) caused an intracellular acidification that was rapidly reversed by Na replacement and occurred independently of Clo. Amiloride (10(-3) M) reversibly blocked Na-dependent recovery after Na-free-induced acidification. These results demonstrate the presence of a Na+-H+ exchange mechanism in pancreatic duct cells. Replacement of Clo with gluconate caused an intracellular alkalinization that was reversed by replacement of Cl. Application of the disulfonic stilbene derivatives, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and dihydro-DIDS (2 X 10(-4) M), resulted in block of both Cl-dependent recovery from Na-gluconate and the onset of alkalinization of transition from NaCl to Na-gluconate. Chloride-dependent alteration of pHi occurred independently of Nao. These results demonstrate the presence of an anion exchange mechanism consistent with Cl--HCO3- exchange. Thus pancreatic duct cells contain both Na+-H+ and Cl--HCO3- exchangers.

Acid-Base Equilibrium↗

PubMed2006

Extracellular NAD+ metabolism modulates osteoclastogenesis.

Changes in NAD+ and its metabolites contribute to longevity and age-associated diseases. The role of NAD+ metabolism in bone diseases has however not been investigated, despite the fact that osteoporosis is a leading cause of morbidity in old age. TRAP(+) osteoclast formation from C57 Bl/6J mice was assessed after the addition of varying concentrations of NAD+ metabolites or exogenous ADPribosyl cyclase and NADase enzymes. The NAD+ metabolite cyclic ADPribose (cADPr) or exogenous addition of the enzyme ADPribosyl cyclase stimulated osteoclast formation. Blocking cADPr action with the antagonist 8-Br-cADPr potently inhibited osteoclast formation. In contrast to cADPr, its noncyclized derivative ADPribose (ADPr) or the exogenous addition of NADase both inhibited osteoclastogenesis. As CD38 is the major NAD+ -degrading enzyme present in the bone marrow, these results suggest that CD38-mediated inhibition of osteoclastogenesis is related to its NADase activity, not its ADPribosyl cyclase activity.

ADP-ribosyl Cyclase 1↗

PubMed2002

Vanadate trapping of nucleotide at the ATP-binding sites of human multidrug resistance P-glycoprotein exposes different residues to the drug-binding site.

The human multidrug resistance P-glycoprotein uses ATP to transport a wide variety of structurally unrelated cytotoxic compounds out of the cell. In this study, we used cysteine-scanning mutagenesis and cross-linking studies to identify residues that are exposed to the drug-binding site upon vanadate trapping. In the absence of nucleotides, C222(TM4) was cross-linked to C868(TM10) and C872(TM10); C306(TM5) was cross-linked to C868(TM10), C872(TM10), C945(TM11), C982(TM12), and C984(TM12); and C339(TM6) was cross-linked to C868(TM10), C872(TM10), C942(TM11), C982(TM12), and C985(TM12). These cysteines are in the middle of the predicted transmembrane (TM) segments and form the drug-binding site. Cross-linking between 332C(TM6) and cysteines introduced at the extracellular side of other TM segments was also done. In the absence of nucleotides, residues 332C and 856C on the extracellular side of TMs 6 and 10, respectively, were cross-linked with a 13-A cross-linker (M8M, 3,6-dioxaoctane-1,8-diyl bismethanethiosulfonate). ATP plus vanadate inhibited cross-linking between 332C(TM6) and 856C(TM10) as well as those in the drug-binding site. Instead, vanadate trapping promoted cross-linking between 332C(TM6) and 976C(TM12) with a 10-A cross-linker (M6M, 1,6-hexanediyl bismethanethiosulfonate). When ATP hydrolysis was allowed to proceed, then 332C(TM12) could form a disulfide bond with 975C(TM12). The cross-linking pattern of 332C(TM6) with residues in TM10 and TM12 indicates that the drug-binding site undergoes dynamic and relatively large conformational changes, and that different residues are exposed to the drug-binding site during the resting phase, upon vanadate trapping and at the completion of the catalytic cycle.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2006

Release of secondary free radicals during post-ischaemic reperfusion is not influenced by extracellular calcium levels in isolated rat hearts.

In this study, we evaluated the impact of the calcium concentration present in the perfusion medium (1.2-3 mM) on contractile performance, lactate dehydrogenase (LDH) release and secondary free radical production during post-ischaemic reperfusion of isolated rat hearts. The impact of calcium concentration on post-ischaemic free radical release was investigated using the Electron Paramagnetic Resonance (EPR) technique and spin trapping with the lipophilic spin trap alpha-phenyl N-tert-butylnitrone (PBN). The evolution of left ventricular end diastolic pressure (LVEDP) in both groups followed the same pattern, but we observed that ischaemic and post-ischaemic contracture was more severe in the group of hearts perfused with 3 mM of calcium as compared with those perfused with 1.2 mM of calcium. A large release of alkyl/alkoxyl species occurred in all hearts from the onset of reperfusion and remained at a high level during the 30 min of reperfusion with no return to basal values. The kinetics and intensity of these releases were the same in both groups. In conclusion, in a range of extracellular calcium levels (1.2-3 mM), the release of alkyl/alkoxyls radicals does not seem to be calcium-dependent. Due to the protective actions of PBN itself, the results of simultaneous investigations of the effects of radical scavengers on isolated heart function may be limited. However, since many pharmacological properties (antioxidant, cellular protector, NO precursor ...) are attributed to PBN, studies investigating oxidative stress with such a multi-faceted tool make interpretation difficult.

Animals↗

PubMed1984

The influence of electrochemical gradients of Na+ and K+ upon the membrane binding and pore forming activity of the terminal complement proteins.

The hemolytic activity of the terminal complement proteins (C5b-9) towards erythrocytes containing high potassium concentration has been reported to be dramatically increased when extracellular Na+ is substituted isotonically by K+ (Dalmasso, A.P., et al., 1975, J. Immunol. 115:63-68). This phenomenon was now further investigated using resealed human erythrocyte ghosts (ghosts), which can be maintained at a nonlytic osmotic steady state subsequent to C5b-9 binding: (1) The functional state of C5b-9-treated ghosts was studied from their ability to retain trapped [14C]-sucrose or [3H]-inulin when suspended either in the presence of Na+ or K+. A dramatic increase in the permeability of the ghost membrane to both nonelectrolytes - in the absence of significant hemoglobin release - was observed for C5b-9 assembly in the presence of external K+. (2) The physical binding of the individual 125I-labeled terminal complement proteins to ghost membranes was directly measured as a function of intra- and extracellular K+ and Na+. The uptake of 125I-C7, 125I-C8, and 125I-C9 into membrane C5b-9 was unaltered by substitution of Na+ by K+. (3) The binding of the terminal complement proteins to ghosts subjected to a transient membrane potential generated by the K+-ionophore valinomycin (in the presence of K+ concentration gradients) was measured. No significant change in membrane binding of any of the C5b-9 proteins was detected under the influence of both depolarizing and hyperpolarizing membrane potentials.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Membrane Permeability↗

PubMed1994

Hormone-binding properties and glycosylation pattern of a recombinant form of the extracellular domain of the luteinizing hormone/chorionic gonadotropin receptor expressed in mammalian cells.

The present studies were undertaken to characterize the properties of a recombinant truncated rat LH/CG (rLH/CG) receptor representing the extracellular domain (rLHR-t338) expressed in mammalian cells. Using detergent-soluble extracts of stably transfected cells, we show that the binding of ovine (o) LH to the extracellular domain is different from the binding of oLH to the full-length receptor. Thus, whereas the full-length receptor from either rat corpora lutea or stably transfected 293 cells binds oLH with two apparent affinities, rLHR-t338 binds oLH with only one affinity, which is intermediate between the high and low affinities of the full-length receptors. However, rLHR-t338 as well as the full-length receptor binds hCG and hLH with similar high affinities and by a model assuming the presence of only one binding site. These data are consistent with the possibility that the carboxyl-terminal half of the LH/CG receptor may contribute (either directly or indirectly) to a second low affinity hormone-binding site. The truncated receptor retains normal binding specificity, as it does not bind hFSH, demonstrating that the extracellular domain alone contains all of the necessary elements for conferring specificity among the gonadotropin hormones. Our studies also show that rLHR-t338 is glycosylated and migrates as two proteins of 48 and 44 kilodaltons (kDa) on sodium dodecyl sulfate gels (where the 44-kDa species is probably a partially degraded form of the 48-kDa protein). However, the N-linked oligosaccharides of the truncated receptor do not appear to be fully processed, as they remain in a high mannose form. rLHR-t338 is not secreted from cells, but remains trapped intracellularly. It can be detected, though, in detergent-soluble extracts of cells or in homogenates. Upon centrifugation of homogenates, it was found that about 90% of the recoverable truncated receptor can be obtained in the supernatant fraction. Thus, mammalian cells could provide a source of a water-soluble extracellular domain of the LH/CG receptor.

Animals↗

PubMed1985

K+-depolarization induces a direct release of Ca2+ from intracellular storage sites in cultured vascular smooth muscle cells from rat aorta.

Using an intracellularly trapped dye, quin 2, effects of K+-depolarization on cytosolic free calcium concentrations were recorded microfluorometrically in rat aorta vascular smooth muscle cells in primary culture. When the cells were exposed to high extracellular K+ in Ca+-free media containing 2mM EGTA, there was a transient and dose-dependent elevation of cytosolic Ca2+ concentrations. However, the concentration of the cytosolic Ca2+ was not elevated when the intracellularly stored Ca2+ was depleted by the repetitive treatment with caffeine prior to the application of high K+. Thus depolarization of plasma membrane, per se, directly induces a release of Ca2+ from intracellular storage sites in vascular smooth muscle cells, and the main fraction of this released Ca2+ is derived from the caffeine sensitive storage sites; perhaps from the sarcoplasmic reticulum.

Aminoquinolines↗

PubMed1997

Cardiac microdialysis of salicylic acid .OH generation on nonenzymatic oxidation by norepinephrine in rat heart.

The effect of pargyline, a monoamine oxidase inhibitor, on the generation of hydroxyl free radicals (.OH) was investigated using cardiac microdialysis. Salicylic acid in Ringer's solution (0.5 nmol x microL(-1) x min(-1)) was infused directly through a microdialysis probe to detect the generation of .OH as reflected by the formation of dihydroxybenzoic acid (DHBA) in the myocardium of anesthetized rats. When pargyline (100 nmol x microL(-1) x min(-1)) was infused in rat heart, the level of norepinephrine (NE) gradually increased in a time-dependent manner and an increase of DHBA was also observed. When NE was administered to the pargyline pretreated animals, a marked elevation in the levels of 2,3- and 2,5-DHBA formation was obtained, as compared to the group treated with NE only, showing a positive linear correlation between NE and .OH formation trapped as 2,3-DHBA (R2 = 0.981) or 2,5-DHBA (R2 = 0.984) in the dialysate. NE clearly produced an increase in .OH formation. These results indicate that accumulation of NE in the extracellular fluid elicited by pargyline can be auto-oxidized, which in turn, leads (possibly by an indirect mechanism) to the formation of cytotoxic .OH free radicals.

Animals↗

PubMed2003

Role of glypican-1 in the trophic activity on PC12 cells induced by cultured sciatic nerve conditioned medium: identification of a glypican-1-neuregulin complex.

Glypican-1 is an extracellular matrix component found by microsequencing in a medium conditioned by cultured rat-sciatic nerves (CM). This CM was concentrated by ultrafiltration and fractionated by quaternary ammonium chromatography, followed by Hi-Trap blue affinity chromatography to obtain the active fraction B1.2. Previously, we have reported a 54 kDa neuregulin (NRG) in the same B1.2 fraction [Villegas et al., Brain Res. 852 (2001) 304]. The effect of Glypican-1 on the neuron-like differentiation of PC12 cells was investigated by immunoprecipitation, Western blot and cellular image analysis. Removal of glypican-1 by immunoprecipitation with increasing concentrations of specific antibodies revealed a gradual decrease of the differentiation activity of fraction B1.2, which paralleled the results obtained by removal of the 54 kDa NRG protein. Colorless native electrophoresis and Western blot analysis was used to identify a glypican-1-NRG protein complex, which could be afterwards separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis into its individual components. Additionally, it was demonstrated that glypican-1, in cooperation with the 54 kDa NRG, is involved in the neuronal-like differentiation of PC12 cells and could play an important role on the regeneration responses of peripheral nerves.

Animals↗