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1.
Isoelectric focusing was performed on parotid salivas selected for their electrophoretic phenotypes of proline-rich acidic salivary proteins. Fractions encompassing narrow pH regions were pooled and examined by polyacrylamide gel electrophoresis. Isoelectric focusing yielded partial purification of major and minor acidic proline-rich proteins which were subsequently compared by immunoelectrophoresis and double immunodiffusion against goat anti-human parotid saliva. Cross-reactivity without spurring between all fractions containing major Pr proteins in both immunoelectrophoresis and double immunodiffusion suggests that these proteins are immunologically very similar or identical.This study was supported in part by an award from the American Cancer Society Institutional Grant IN-88F to Fels Research Institute. 相似文献
2.
Sheep erythrocytes (E) which, with or without certain treatments, are currently used as “immunological reagents” to detect
cells with specific receptors (by rosette-formation) have been partitioned in two-polymer aqueousphase systems selected so
as to reflect charge-associatedor lipid-related membrane surface properties. We have found that the partitioning behavior of E is not affected in these phases
by reacting the cells with anti-E antibody (either IgG or IgM), forming EA. The additional binding of complement to the cell-antibody
complex, forming EAC, results, however, in a marked decrease in the partition coefficient,K. Apparently both the charge-associated and hydrophobic properties reflected by partitioning remain accessible to the phase
polymers when the cells are coated with antibody, but are not with the addition of complement. It is interesting that EA can
still rosette with T-lymphocytes (14), a property of E, while the additional coating with complement results in EAC which does not appreciably do so (26).
Neuraminidase or trypsin treatments of E, which yield Es having quite different rosetting properties with T-lymphocytes (14), cause increasedKs and unchangedKs, respectively, in phases reflecting lipid-related surface properties. Either treatment causes reducedKs of E in charged-phase systems. Neuraminidase treatment also results in a reduced electrophoretic mobility of E, while trypsin
treatment is not detectable by cell electrophoresis (25). We are currently studying the possible usefulness of employing cell electrophoresis and cell partitioning in charged-phase
systems jointly to obtain information on events occurring at the shear plane versus those occurring deeper in the membrane. 相似文献
3.
4.
An early event in the formation of the serotonergic synapse by the Retzius (R) onto the pressure-sensitive (P) neurons of the leech is the elimination of an extrasynaptic response to transmitter from sites of contact on the postsynaptic cell. This event during synapse formation is cell-specific in that it is elicited in vitro by contact with the presynaptic R cell but not with other neurons. In the study reported here, we investigated the nature of this interaction between R and P neurons. The loss of the extrasynaptic response of the P cell was elicited by contact with R cells fixed in a mild paraformaldehyde solution, but not by R cells treated with the proteolytic enzyme trypsin prior to fixation. As well, a variety of lectins were assayed for their ability to interfere with synapse formation. The transmitter responses of P cells plated on lectin-coated substrates were unaffected. However, exposure of the R cell to the lectin wheat germ agglutinin (WGA), but not to other lectins, prior to pairing prevented the loss of the extrasynaptic response in contacted P cells and blocked the formation of the R? P synapse in culture. We conclude that recognition by the P cell of the R cell during synapse formation may be mediated by an R cell-specific surface protein which binds wheat germ agglutinin. 1994 John Wiley & Sons, Inc. 相似文献
5.
To study the gene expression profiles between immunologically injured liver cell and normal liver cell of mice and to screen
on a large scale the differentially expressed genes associated with the formation of liver injury, the experimental mice were
randomly divided into the normal group for controlling and the immunologically liver-injured group induced by BCG and LPS.
The liver mRNA of the two groups were extracted respectively and reversely-transcribed to cDNA with the incorporation of different
fluorescence (Cy3, Cy5) labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray
chips. The fluorescent signal results were acquired by scanner ScanArray 4000 and analyzed with software GenePix Pro 3.0.
Among the 14112 target genes, 293 genes were found to be significantly differentially expressed, in which 188 genes were up-regulated
and 105 genes were down-regulated. Based on the analysis of biological functions of those differentially expressed genes,
it was indicated that the occurrence and development of mouse liver damage induced by BCG and LPS were highly correlated with
the processes of immune reactions, cell synthesis, metabolism, apoptosis and transportation in liver cell, which might be
quite important for elucidating the regulatory network of gene expression associated with the liver damage, also important
for finally discovering the pathogenic mechanisms of immunological liver damage. 相似文献
6.
The direct visualization of subcellular dynamic processes is often hampered by limitations in the resolving power achievable with conventional microscopy techniques. Fluorescence recovery after photobleaching has emerged as a highly informative approach to address this challenge, permitting the quantitative measurement of the movement of small organelles and proteins in living functioning cells, and offering detailed insights into fundamental cellular phenomena of physiological importance. In recent years, its implementation has benefited from the increasing availability of confocal microscopy systems and of powerful labeling techniques based on genetically encoded fluorescent proteins or other chemical markers. In this review, we present fluorescence recovery after photobleaching and related techniques in the context of contemporary neurobiological research and discuss quantitative and semi‐quantitative approaches to their interpretation. 相似文献
7.
Uchino S Wada H Honda S Nakamura Y Ondo Y Uchiyama T Tsutsumi M Suzuki E Hirasawa T Kohsaka S 《Journal of neurochemistry》2006,97(4):1203-1214
A class of scaffolding protein containing the post-synaptic density-95/Dlg/ZO-1 (PDZ) domain is thought to be involved in synaptic trafficking of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors during development. To clarify the molecular mechanism of AMPA receptor trafficking, we performed a yeast two-hybrid screening system using the cytoplasmic tail of the GluR1 subunit of AMPA receptor as a bait and identified a synaptic molecule, Shank3/ProSAP2, as a GluR1 subunit-interacting molecule. Shank3 is a PDZ domain-containing multidomain protein and is predominantly expressed in developing neurons. Using the glutathione S-transferase pull-down assay and immunoprecipitation technique we demonstrated that the GluR1 subunit directly binds to the PDZ domain of Shank3 via its carboxyl terminal PDZ-binding motif. We raised anti-Shank3 antibody to investigate the expression of Shank3 in cortical neurons. The pattern of Shank3 immunoreactivity was strikingly punctate, mainly observed in the spines, and closely matched the pattern of post-synaptic density-95 immunoreactivity, indicating that Shank3 is colocalized with post-synaptic density-95 in the same spines. When Shank3 and the GluR1 subunit were overexpressed in primary cortical neurons, they were also colocalized in the spines. Taken together with the biochemical interaction of Shank3 with the GluR1 subunit, these results suggest that Shank3 is an important molecule that interacts with GluR1 AMPA receptor at synaptic sites of developing neurons. 相似文献
8.
9.
Alves Luís Cyrne Luisa Amaral-Collaço M.T. Gírio Francisco M. 《World journal of microbiology & biotechnology》2003,19(2):201-208
Antisera against metal(Mo)-containing dye-linked dehydrogenases from sulphate-reducing bacteria were used to screen for immunological similarities with NAD+-linked dehydrogenases detected in aerobic methanol-utilizing bacterial isolates. Out of eleven strains tested, the strains #5, 8, 9 and 11 were shown to have specific formate and aldehyde dehydrogenases displaying antibody cross-reaction against highly purified Mo-containing dye-linked dehydrogenases. The apparent molecular mass of the identified proteins observed during the antibody reaction correlated with the molecular mass of the dehydrogenases obtained after native PAGE electrophoresis. The strains #8 and 11 exhibited one formate dehydrogenase apparently of identical molecular mass 140–145 kDa, whereas strains #5, 9 and 11 synthesized aldehyde dehydrogenases with apparent molecular masses of about 110, 120 and 155 kDa (two forms) and 120 kDa, respectively. All these aerobic enzymes shared antigenic properties with the anaerobic metalloproteins, indicating the existence of structural similarities between those enzymes in spite of having different cofactor moieties. 相似文献
10.
Mathew P. Daniels 《Molecular neurobiology》1997,14(3):143-170
Reciprocal signals between the motor axon and myofiber induce structural and functional differentiation in the developing
neuromuscular junction (NMJ). Elevation of presynaptic acetylcholine (ACh) release on nerve-muscle contact and the correlated
increase in axonal-free calcium are triggered by unidentified membrane molecules. Restriction of axon growth to the developing
NMJ and formation of active zones for ACh release in the presynaptic terminal may be induced by molecules in the synaptic
basal lamina, such as S-laminin, heparin binding growth factors, and agrin. Acetylcholine receptor (AChR) synthesis by muscle
cells may be increased by calcitonin gene-related peptide (CGRP), ascorbic acid, and AChR-inducing activity (ARIA)/heregulin,
which is the best-established regulator. Heparin binding growth factors, proteases, adhesion molecules, and agrin all may
be involved in the induction of AChR redistribution to form postsynaptic-like aggregates. However, the strongest case has
been made for agrin's involvement. “Knockout” experiments have implicated agrin as a primary anterograde signal for postsynaptic
differentiation and muscle-specific kinase (MuSK), as a putative agrin receptor. It is likely that both presynaptic and postsynaptic
differentiation are induced by multiple molecular signals. Future research should reveal the physiological roles of different
molecules, their interactions, and the identity of other molecular participants. 相似文献