SEARCH · PubMed Health

Results for “Chain Migration”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 307 records · Page 17Linked to original sources

PubMed1989

The distribution of heavy-chain isoforms of myosin in airways smooth muscle from adult and neonate humans.

Changes in the expression of heavy chains of myosin during development determine the functional characteristics of striated muscles. The distribution of heavy-chain isoforms of smooth-muscle myosin was determined in the airways of adult and infant humans to see whether it might underlie the hyperreactivity of human airways. The protein bands corresponding to myosin were separated using SDS/polyacrylamide-gel electrophoresis (4% gels) and identified by immunoblotting using both monoclonal and polyclonal antibodies against smooth-muscle myosin and non-muscle myosin. The relative proportion of each heavy chain stained by Coomassie Blue was measured by densitometric scanning. Three major bands corresponding to myosin heavy-chain isoforms were found; the two slower migrating bands (MHC1 and MHC2) were smooth-muscle myosin, and the third band was non-muscle myosin. The MHC1/MHC2 ratio was 0.69:1 in adult bronchus, and in infant bronchus and trachea. This contrasted with the airway smooth muscle in pigs, which was run concurrently, where the smooth-muscle heavy-chain ratio changed with development [Mohammad & Sparrow (1988) FEBS Lett. 228, 109-112]. The non-muscle myosin heavy chain comprised 63% of the smooth-muscle myosin. In both adult and infant lungs an additional putative myosin heavy chain which migrated slightly more rapidly than non-muscle myosin heavy chain was identified using the monoclonal smooth-muscle myosin antibody BF 48. This was unique to the human species.

Aged↗

PubMed1993

Primary structure of Xenopus laevis S10, a ribosomal protein that cross-reacts with antibodies to immunoglobulin light chains.

Screening of a cDNA expression library from Xenopus laevis splenocytes with purified antibodies to Xenopus immunoglobulin light chains unexpectedly led to the isolation of a clone with an insert whose deduced amino acid sequence is similar to that of a segment of a protein, S10, from the small (40S) subunit of rat ribosomes. A clone containing an insert encoding the corresponding complete protein was isolated from another cDNA library by nucleic acid hybridization. The deduced amino acid sequence of this insert is 94% identical to that of rat S10; no similarity to immunoglobulin sequences could be discerned. The reactivity of the anti-light chain antibodies with the putative Xenopus S10 facilitated the purification of the protein, by high-pressure liquid chromatography, from the 40S subunit of Xenopus ribosomes. Amino-terminal sequence analysis established the identity of the ribosomal protein with the protein encoded by the cDNA insert. To explore the basis for this unexpected cross-reaction, an "antibody transfer" experiment was carried out. Antibodies to Xenopus light chains were adsorbed to Xenopus S10 on a nitrocellulose strip, which was incubated with another strip containing separated heavy and light chains from Xenopus IgM. Antibodies migrated from the strip carrying S10 to the light chains, but not the heavy chains, on the second strip. These results suggest that this unexpected cross-reaction is due to the sharing of one or more epitopes by Xenopus immunoglobulin light chains and the ribosomal protein, S10.

Amino Acid Sequence↗

PubMed2005

An investigation into the excitation migration in polyfluorene solutions via temperature dependent fluorescence anisotropy.

Fluorescence anisotropy of dilute polyfluorene polymer solutions has been used to show that two processes, exciton migration and conformational relaxation (twisting of part of the chain), occur within polyfluorene polymers of 20 or more repeat units. The former process is dominant and temperature independent for high excitation energy but, as the chain length is decreased, exciton migration is eliminated and the conformational relaxation becomes the only mechanism by which excited state energy relaxation to the emission site can occur. When the polymers are excited in the absorption band tails, again no migration is observed but conformational relaxation is still present.

Journal Article↗

PubMed1975

[Chemical properties of plasma fibrinogens and fibrins from normal and cobalt-treated rabbits].

The sulfitolysis products of fibrinogens from normal and cobalt-treated rabbits (5 mg Co2+/kg b.w.) were resolved by ion exchange chromatography on CM-cellulose columns. The elution patterns of both fibrinogens showed a distinct heterogeneity of gamma-chains. Furthermore, a gamma-chain derivative from cobalt fibrinogen could be distinguished electrophoretically from the corresponding one of normal fibrinogen because of its reduced electrophoretic mobility. Normal and cobalt fibrinogen did not differ from each other in their N-terminal (Val2 Ala2 Tyr2) and C-terminal (Pro1-2 Val4-5) amino acid compositions related to a subunit structure of A alpha 2 B beta 2 gamma 2, and their carbohydrate contents - neutral hexoses 1,21% (1,26%), N-acetyl hexosamines 1,16% (1,05%), N-acetyl neuraminic acid 1,19% (1, 13%), values for cobalt fibrinogen in parentheses. The main amounts of carbohydrates are bound to gamma- and B beta-chains, The BrCn cleavage products from cobalt fibrinogen and its gamma- and B beta-chains showed other electrophoretic properties than the corresponding derivatives from normal fibrinogen. But BrCN split products of A alpha-chains of both fibrinogens were electrophoretically very similar. Spectrographic investigations of the S-sulfoderivates demonstrated a diminution of the absorption maximum near 282mn of gamma and B beta derivatives of cobalt fibrinogen. A alpha-chains of both fibrinogens were not different from each other. Using autoradiography the highest 58Co binding could be found in the gamma-chain with a reduced electrophoretic migration velocity, whereas B beta-and gamma-chains with unchanged electrophoretic mobility bound only small amounts. A alpha-chains of cobalt fibrinogens were apparently not loaded with 58Co. gamma-chain and alpha-chain cross-links could be observed in normal fibrins stabilized by factor XII, however, in cobalt fibrins a gamma-dimer formation was demonstrable without participation of gamma-chains with reduced electrophoretic mobility. A distinct alpha-chain cross-link could not be demonstrated either. From these and other investigations on molecular weights of both fibrinogens it was assumed that earlier observed changes of physicochemical properties and biological behaviour of cobalt fibrinogen might result from a complex binding of cobalt ions on specific structures of the fibrinogen molecule.

Amino Acid Sequence↗

PubMed1996

Network of tangential pathways for neuronal migration in adult mammalian brain.

Cells in the brains of adult mammals continue to proliferate in the subventricular zone (SVZ) throughout the lateral wall of the lateral ventricle. Here we show, using whole mount dissections of this wall from adult mice, that the SVZ is organized as an extensive network of chains of neuronal precursors. These chains are immunopositive to the polysialylated form of NCAM, a molecule present at sites of plasticity, and TuJ1, an early neuronal marker. The majority of the chains are oriented along the rostrocaudal axis and many join the rostral migratory stream that terminates in the olfactory bulb. Using focal microinjections of DII and transplantation of SVZ cells carrying a neuron-specific reporter gene, we demonstrate that cells originating at different rostrocaudal levels of the SVZ migrate rostrally and reach the olfactory bulb where they differentiate into neurons. Our results reveal an extensive network of pathways for the tangential chain migration of neuronal precursors throughout the lateral wall of the lateral ventricle in the adult mammalian brain.

Animals↗

PubMed2002

Thymosin beta4 promotes human conjunctival epithelial cell migration.

PURPOSE: In this study the effects of thymosin beta4 (Tbeta4) on migration and production of laminin-5 in the human conjunctival cell line HC0597 was analyzed. METHODS: Boyden chamber assays assessed the ability of Tbeta4 to stimulate in vitro cell migration. Control or Tbeta4-treated cells were processed for immunofluorescence microscopy using antibodies to vinculin or laminin-5. Cell lysates were processed for Western blot and densitometric analysis using antibodies to laminin-5 alpha3 or gamma2 chains. RESULTS: Tbeta4 stimulated migration in a dose-dependent manner. Focal adhesions present in Tbeta4-treated cells were smaller and more rounded compared to the "streaks" characteristic of controls. Western blot analysis and densitometry revealed that Tbeta4-treated cells expressed more laminin-5 alpha3 and gamma2 chain protein. CONCLUSION: Tbeta4 stimulates in vitro conjunctival epithelial cell migration, and results in altered focal adhesion formation and increased extracellular laminin-5 deposition. The increased migration may be correlated with increased production of laminin-5.

Blotting, Western↗

PubMed1996

Purification, characterization and partial amino acid sequencing of two new aspartic proteinases from fresh flowers of Cynara cardunculus L.

Two new aspartic proteinases have been isolated from stigmas of the cardoon Cynara cardunculus L. by a two-step purification procedure including extraction at low pH, gel filtration on Superdex 200, and ion-exchange chromatography on Mono Q. To follow the conventional nomenclature for aspartic proteinases, we have named these proteinases cardosin A and cardosin B. On SDS/PAGE, cardosin A migrated as two bands with apparent molecular masses of 31 000 Da and 15 000 Da whereas the chains of cardosin B migrated as bands of 34 000 Da and 14 000 Da. The partial amino acid sequences of the two cardosin revealed that they are similar but not identical, and that they differ from the previously reported cardoon proteinases named cynarases, which were assumed to be derived from a common precursor. Although the cardosins show some degree of similarity to each other, we could detect no immunological crossreactivity between them. Both cardosins were active at low pH and were inhibited by pepstatin, with Ki values of 3 nM for cardosin A and 1 nM for cardosin B, indicating that they belong to the class of aspartic proteinases. Significant differences between the two enzymes were also found for the Kcat/km values for the hydrolysis of two chromophoric synthetic peptides. The active-site ionization constants, pKe1 and pKe2, for cardosin A are 2.5 +/- 0.2 and 5.3+/- 0.2, whereas for cardosin B they are 3.73 +/- 0.09 and 6.7 +/- 0.1. The results herein described on the structural and kinetic properties of the cardosins indicate that they are the products of distinct genes which have probably arisen by gene duplication. A scheme for the proteolytic processing of the two enzymes is also proposed.

Amino Acid Sequence↗

PubMed1993

VLA-4 integrin can mediate CD11/CD18-independent transendothelial migration of human monocytes.

The migration of human monocytes across unactivated and activated human umbilical vein endothelium (HUVE) in response to chemotactic factors was studied, and the adhesion molecules involved were characterized. Migration of blood monocytes or U937 cell line-derived monocytes across unactivated HUVE induced by C5a, was partially inhibited (by 75%) by mAbs (R15.7 or 60.3) to CD18 of the CD11/CD18 complex on the monocyte. However, when the HUVE was pretreated for 5 h with IL-1 alpha (0.1 ng/ml), TNF-alpha (100 U/ml), or LPS (1 ng/ml), migration induced by C5a was no longer inhibited; i.e., migration became CD18 independent. The monocyte CD18-independent migration was completely blocked by mAbs against alpha 4 or beta 1 integrin chains of VLA-4. This migration was also partially inhibited by mAbs against vascular cell adhesion molecule-1 (VCAM-1), a major counter-receptor on HUVE for VLA-4, but not by mAbs to E-selectin or intercellular adhesion molecule-1. The significant CD18-independent migration across "unactivated" HUVE was also inhibited by mAbs against alpha 4 or beta 1 chains of VLA-4, although mAbs against VCAM-1 did not inhibit under these conditions. Finally, considerable VLA-4-dependent transendothelial migration to C5a was also observed with monocytes from a patient with CD18 deficiency (leukocyte adhesion deficiency). These results suggest that (a) there is a major CD18-independent component in monocyte chemotactic factor-dependent migration across activated and unactivated endothelium; (b) that VLA-4 integrin on the monocyte has a major role in this migration; and (c) that VCAM-1 on activated endothelium functions as a counter-receptor in this process, but other ligands for VLA-4, especially on unactivated endothelium, may also be involved.

Antibodies, Monoclonal↗

PubMed2003

Differential regulation of cellular adhesion and migration by recombinant laminin-5 forms with partial deletion or mutation within the G3 domain of alpha3 chain.

The basement membrane protein laminin-5 promotes cell adhesion and migration. The carboxyl-terminal G3 domain in the alpha3 chain is essential for the unique activity of laminin-5. To investigate the function of the G3 domain, we prepared various recombinant laminin-5 forms with a partially deleted or mutated G3 domain. The deletion of the carboxyl-terminal 28 amino acids (region III) markedly decreased the cell adhesion activity with a slight loss of the cell motility activity toward BRL and EJ-1 cells. This change was attributed to the loss of Lys-Arg-Asp sequence. Further deletion of 83 amino acids (region II) led to almost complete loss of the cell motility activity. All charged amino acid residues tested in this region were not responsible for the activity loss. These results suggest that the G3 domain contains two distinct regions that differently regulate cell adhesion and migration. Analysis of laminin-5 receptors showed that integrins alpha3beta1, alpha6beta1, and alpha6beta4 had different but synergistic effects on cell adhesion and migration on laminin-5. However, the structural change of the G3 domain appeared not to change integrin specificity. The present study demonstrates that the G3 domain in laminin-5 plays a central role to produce different biological effects on cells.

Amino Acid Sequence↗

PubMed1991

Myosin heavy chain isoforms in white, red and ventricle muscles of barbel (Barbus barbus L.).

1. Actomyosin extracts of trunk, heart, and head muscles from barbel (Barbus barbus L.) were analyzed by SDS-polyacrylamide gel electrophoresis to study their myosin heavy chain composition. 2. Four heavy chain isoforms were found: trunk white, trunk red, and ventricle muscles yielded one heavy chain typical of the muscle type; head muscles devoid of red fibers displayed two heavy chain isoforms, the slow migrating one corresponding to the trunk white muscle type. 3. The electrophoretic mobility of red and ventricle myosin heavy chains related to that of white isoforms appeared highly modified by the glycerol content of the gels.

Animals↗

PubMed2001

Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris.

A custom-designed, highly hydrophilic gelatin was produced in Pichia pastoris. Secreted production levels in single-copy transformants were in the range 3-6 g/l of clarified broth and purification to near homogeneity could be accomplished by differential ammonium sulfate precipitation. Despite the fact that gelatins are highly susceptible to proteolysis because of their unfolded structure, the recombinant protein was shown to be fully intact by SDS-PAGE, N-terminal sequencing, gel filtration chromatography and mass spectrometry. Owing to its highly hydrophilic nature, the migration of the synthetic gelatin in SDS-PAGE was severely delayed. Esterification of the carboxylic amino acid side chains resulted in normal migration. The high polarity of the synthetic gelatin also accounts for its negligible surface activity in water at concentrations up to 5% (w/v), as determined by tensiometry. Circular dichroism spectrometry showed that the non-hydroxylated gelatin did not form triple helices at 4 degrees C. The spectrum was even more representative of the random coil conformation than the spectrum of natural non-hydroxylated gelatins.

Base Sequence↗

PubMed1987

Spectrin Nice (beta 220/216): a shortened beta-chain variant associated with an increase of the alpha I/74 fragment in a case of elliptocytosis.

We describe a new spectrin variant with a truncated beta-chain. It was discovered in a 17-year-old white boy presenting with intermittent jaundice and spleen enlargement. He also displayed numerous smooth elliptocytes. On sodium dodecyl sulfate-polyacrylamide gel, the truncated beta-chain (beta'-chain) appeared as an additional band of approximately 216 kilodaltons, migrating between spectrin beta-chain and ankyrin. It represented 30% of total beta-chain. The beta'-chain reacted with an antispectrin beta-chain monoclonal antibody. It failed to become phosphorylated when ghosts were incubated in the presence of [gamma-32P] adenosine triphosphate. Whole spectrin tetramerization was defective since the amount of spectrin dimer was increased in spectrin crude extract and the association constant of the spectrin dimer self-association was decreased. Spectrin whole tetramer isolated from spectrin crude extracts contained small quantities of beta'-chain. Spectrin tryptic peptides showed an increase of the 74,000-dalton fragment at the expense of the 80,000-dalton fragment. So far, the latter abnormality has been used to characterize a number of cases of hereditary elliptocytosis or pyropoikilocytosis with no other apparent change. In the present case, we consider that the abnormality is a consequence of the beta-chain alteration. The parents seemed asymptomatic. As a result, we regard this new spectrin variant as deriving from a de novo mutation.

Adolescent↗

PubMed1997

Direct evidence for homotypic, glia-independent neuronal migration.

Neuronal precursors born in the subventricular zone (SVZ) of the neonatal and adult rodent brain migrate 3-8 mm from the walls of the lateral ventricle into the olfactory bulb. This tangentially oriented migration occurs without the guidance of radial glia or axonal processes. The cells move closely associated, forming elongated aggregates called chains, which are ensheathed by astrocytes. We have developed a culture system in which postnatal mouse SVZ neuronal precursors assemble into chains with ultrastructural and immunocytochemical characteristics equivalent to those in vivo but without the astrocytic sheath. Time-lapse videomicrography revealed that individual cells migrate along the chains very rapidly (approximately 122 microm/hr) in both directions. Periods of cell body translocation were interspersed with stationary periods. This saltatory behavior was similar to radial glia-guided migration but approximately 4 times faster. Neuronal precursors isolated from embryonic cortical ventricular zone or cerebellar external granule layer did not form chains under these conditions, suggesting that chain migration is characteristic of SVZ precursors. This study directly demonstrates that SVZ neuronal precursors migrate along each other without the assistance of astrocytes or other cell types. (Additional data are presented in www.cell.com).

Animals↗

PubMed1992

Baculovirus expression of mammalian G protein alpha subunits.

Complementary DNAs encoding three subtypes of the alpha subunit (alpha i-1, alpha o and alpha s) of rat guanyl nucleotide regulatory proteins were used to construct recombinant baculoviruses which direct high-level expression of the corresponding proteins in cultured Sf9 insect cells. The expressed proteins were recognized by polyclonal antisera specific for the different alpha chains, and co-migrated with the native proteins from rat brain membranes in immunoblotting analyses. Soluble and particulate forms of all three immunoreactive alpha chains were observed following ultracentrifugation of cell lysates. Biosynthetic radiolabelling of infected cells with [35S]methionine or [3H]myristate showed that both soluble and particulate forms of alpha i-1 and alpha o were myristoylated; in contrast, alpha s did not incorporate myristate. The soluble fractions from cells expressing alpha chains showed high levels of GTP-binding activity over that observed in uninfected cells, or in cells infected with wild-type virus. The peak expression levels observed at 72 h post-infection were highest for alpha o at ca. 400 pmol of GTP-gamma-35S/mg protein, or roughly 2% of the total soluble protein. The results of this work show that the baculovirus system can be employed for high-level production of mammalian G protein alpha chains which retain GTP-binding activity and are appropriately modified by myristoylation.

Animals↗

PubMed1997

Differential localization of myosin and myosin phosphatase subunits in smooth muscle cells and migrating fibroblasts.

Myosin II light chains (MLC20) are phosphorylated by a Ca2+/calmodulin-activated kinase and dephosphorylated by a phosphatase that has been purified as a trimer containing the delta isoform of type 1 catalytic subunit (PP1C delta), a myosin-binding 130-kDa subunit (M130) and a 20-kDa subunit. The distribution of M130 and PP1C as well as myosin II was examined in smooth muscle cells and fibroblasts by immunofluorescence microscopy and immunoblotting after differential extraction. Myosin and M130 colocalized with actin stress fibers in permeabilized cells. However, in nonpermeabilized cells the staining for myosin and M130 was different, with myosin mostly at the periphery of the cell and the M130 appearing diffusely throughout the cytoplasm. Accordingly, most M130 was recovered in a soluble fraction during permeabilization of cells, but the conditions used affected the solubility of both M130 and myosin. The PP1C alpha isoform colocalized with M130 and also was in the nucleus, whereas the PP1C delta isoform was localized prominently in the nucleus and in focal adhesions. In migrating cells, M130 concentrated in the tailing edge and was depleted from the leading half of the cell, where double staining showed myosin II was present. Because the tailing edge of migrating cells is known to contain phosphorylated myosin, inhibition of myosin LC20 phosphatase, probably by phosphorylation of the M130 subunit, may be required for cell migration.

Amino Acid Sequence↗

PubMed1993

Developmental expression of the alpha 6 GABAA receptor subunit mRNA occurs only after cerebellar granule cell migration.

Using a competitive polymerase chain reaction (PCR) and appropriate internal standards, we have analyzed absolute amounts of the alpha 6 GABAA receptor subunit mRNA in the postnatally developing cerebellum and neocortex. The PCR data have shown that absolute amounts of the alpha 6 receptor subunit mRNA in the cerebellum increase dramatically (nearly 100-fold) during the second postnatal week, reaching maximal levels by postnatal day 21 (1 fmol/microgram total RNA). The absolute amount of the alpha 6 GABAA receptor subunit mRNA in the cortex at postnatal day 1 was 2 amol/microgram total RNA and increased to 7 amol/micrograms total RNA by postnatal day 14. No further increase in alpha 6 mRNA expression in the adult cortex was observed. Microscopic analysis of emulsion coated and counterstained sections indicated that alpha 6 GABAA receptor subunit mRNA labeling was only detected in the internal granule cell layer and not in either the external granule cell layer or in migrating granule cells. The alpha 1 GABAA receptor subunit mRNA increased in the cerebellar cortex with a similar temporal profile, although its distribution extended to additional cell types (Purkinje cells, stellate/basket cells and possibly cerebellar astrocytes). The temporal expression of these two GABAA receptor subunit mRNAs is coincident with the formation of synaptic contacts in the granule cell dendrites suggesting that afferent pathways innervating these neurons following cell migration may play a critical role in increasing the expression of mRNAs encoding the alpha 1 and alpha 6 GABAA receptor subunits.

Aging↗

PubMed1996

Enhanced electrophoretic separation and resolution of myosin heavy chains in mammalian and avian skeletal muscles.

We report a sodium dodecyl sulfate-polyacrylamide gel electrophoresis protocol for the reliable separation, with high resolution, of myosin heavy chain isoforms in adult avian (chicken) and mammalian (mouse) skeletal muscles. The sample preparation time can be relatively short, thereby minimizing endogenous proteolytic activity which may otherwise result in dispersed and spurious bands. Inclusion of 2-mercaptoethanol in the upper electrode buffer greatly improves band resolution. Glycerol is commonly included in the reported protocols for myosin heavy chain separation and our results demonstrate that the concentration of glycerol employed can have a marked effect on the relative order of migration among myosin heavy chain isoforms.

Animals↗

PubMed1983

O-antigen conversion in Pseudomonas aeruginosa PAO1 by bacteriophage D3.

The lysogenization of Pseudomonas aeruginosa PAO by phage D3 results in derivatives which are resistant to superinfection by phage D3c by virtue of the fact that homologous phage cannot adsorb to these cells. The serologically and morphologically unrelated phage E79 showed a markedly decreased adsorption rate to the lysogen PAO(D3). Since both of these phages are lipopolysaccharide specific, these results suggested lysogenic conversion of the phage receptor. The lipopolysaccharide was extracted from strain PAO by the hot phenol-water technique, but this procedure was ineffective with PAO(D3). We developed a technique involving cold trichloroacetic acid extraction, followed by ultracentrifugation, digestion of the high-speed pellet with proteinase K, and ultimate purification on CsCl step gradients. The lipopolysaccharide from the wild type had inactivating activity against D3 and E79, whereas that from PAO(D3) inactivated neither. Chromatographic analysis indicated that the convertant lipopolysaccharide was smooth, and quantitative chemical analyses of the two preparations showed no differences in the level of the major fatty acids, amino compounds, or neutral sugars. On the other hand, sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the side chains had a decreased migration rate through the gel matrix. The application of 1H and 13C nuclear magnetic resonance spectroscopic analysis revealed that the PAO side chain is chemically identical to that of serotype O:2a,d, containing 2,3-(1-acetyl-2-methyl-2-imidazolino-5,4)-2,3-dideoxy-D-mannuronic acid, 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, and 2-acetamido-2,6-dideoxy-D-galactose (D-fucosamine). The molecular basis of the conversion event was (i) the introduction of an acetyl group into position 4 of the fucosamine residue and a change in the bonding between trisaccharide repeating units from alpha 1 leads to 4 to beta 1 leads to 4.

Antigens, Bacterial↗