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T Miron

Publications and source records attributed to T Miron.

17 recordsLinked to original sources
PubMed1991

A 25-kD inhibitor of actin polymerization is a low molecular mass heat shock protein.

The 25-kD inhibitor of actin polymerization (25-kD IAP), isolated from turkey smooth muscle (Miron, T., M. Wilchek, and B. Geiger, 1988. Eur. J. Biochem. 178:543-553), is shown here to be a low molecular mass heat shock protein (HSP). Direct sequence analysis of the purified protein, as well as cloning and sequencing of the respective cDNA, disclosed a high degree of homology (67% identity, 80% similarity) to the human 27-kD HSP. Southern blot of chicken genomic DNA disclosed one band, suggesting the presence of a single gene, and Northern blot analysis revealed abundant transcript of approximately 1 kb in gizzard and heart tissues and lower amounts in total 18-d chick embryo RNA and in cultured fibroblasts. Exposure of the latter cells to 45 degrees C resulted in over 15-fold increase in the apparent level of the 25-kD IAP protein, confirming that its expression is regulated by heat shock. Immunofluorescent microscopic localization indicated that after heat treatment, the levels of the 25-kD IAP were markedly increased and the protein was apparently associated with cytoplasmic granules. Heat shock also had a transient, yet prominent, effect on the microfilament system in cultured fibroblasts: stress fibers disintegrated within 10-15 min after incubation at 45 degrees C, yet upon further incubation at the elevated temperature, conspicuous actin bundles were apparently reformed.

Actin Cytoskeleton↗
PubMed1988

Characterization of an inhibitor of actin polymerization in vinculin-rich fraction of turkey gizzard smooth muscle.

We report here on the purification and characterization of a new 25-kDa inhibitor of actin polymerization from turkey gizzard smooth muscle. The protein was purified by chromatography on DEAE-cellulose and hydroxyapatite, as well as by affinity chromatography on an immobilized-antibody column. The purified polypeptide reduced the low-shear viscosity of actin, apparently due to its inhibitory effect on actin polymerization. We demonstrate that this protein is largely responsible for the apparent inhibitory activity previously reported to be associated with smooth muscle vinculin preparations. Three independent monoclonal antibodies prepared against the 25-kDa inhibitor of actin polymerization can effectively adsorb the inhibiting activity of actin polymerization from the crude vinculin preparation or inhibit it. We also show here that the 25-kDa inhibitor of actin polymerization tends to undergo dimerization when maintained in non-reducing buffers, concomitant with the loss of its inhibitory activity. Immunohistochemical labeling of frozen sections, as well as immunoblotting analyzes, indicated that the 25-kDa inhibitor of actin polymerization is particularly enriched in smooth muscle cells and that its distribution is apparently homogenous throughout the cytoplasm showing no apparent enrichment in the vinculin-rich dense plaques located along the endofacial surface of the plasma membrane.

Actins↗
PubMed1987

Partial characterization of a peptidoglycan-protein complex from Fusobacterium nucleatum Fev1.

A protein in the cell wall of Fusobacterium nucleatum Fev1 remained associated with the peptidoglycan during extraction with various detergents and organic solvents. On digestion of this peptidoglycan-protein complex (PPC) with murein hydrolases, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed polypeptide bands with apparent molecular weights (MWs) in the range of 3000 to 40,000. After reaction with maleic anhydride the electrophoretic mobilities of these polypeptide bands increased to those of MWs 3000 to 12,000. The PPC protein showed a limited susceptibility toward trypsin, giving polypeptides that migrated in SDS-PAGE as a diffuse band with MW in the range of 3000 to 6000. The amino acid composition of all polypeptide bands eluted from SDS-PAGE was very similar, whichever enzyme was used for the solubilization of the PPC, and was nearly identical to that found for the protein moiety of the PPC. On the basis of a MW of 3000 for a protein unit, about one molecule of protein was found per five peptidoglycan subunits. Lanthionine was not found associated with released polypeptide, and muramic acid and glucosamine were either absent or present in amounts less than one molecule per protein unit. The PPC was immunogenic in rabbits, and purified anti-PPC IgG reacted with murein hydrolase-released protein separated on SDS-PAGE but preferentially with bands of MWs greater than 18,000.

Bacterial Proteins↗
PubMed1982

Composition of peptidoglycans in Bacteroidaceae: determination and distribution of lanthionine.

Peptidoglycans of organisms belonging to the strictly anaerobic family Bacteroidaceae were investigated for the presence of lanthionine. Different procedures for the quantitation of lanthionine were compared. Performic acid and peroxide oxidation procedures on 35S-labeled peptidoglycan from Fusobacterium nucleatum Fev1 resulted in low yields of cysteic acid (42 and 60%, respectively) and many other additional unidentified oxidation products. Lanthionine was, however, recovered in high yield (89% or more) from acid hydrolysates of unoxidized peptidoglycans. Lanthionine was found exclusively in some species of Fusobacterium, in particular F. nucleatum, F. necrophorum, F. russi, and F. gonidiaformans, for which lanthionine may be ascribed a function as a taxonomic marker. Peptidoglycans of these bacteria are thus proposed to belong to a new chemotype, assigned A1 delta. One strain of Fusobacterium, F. mortiferum VPI 0473 contained both lanthionine and diaminopimelic acid in about equal proportions. Species of F.plauti had a composition atypic of gram-negative cells. Chemotypic differences were also indicated among the species of Bacteroides investigated. Thus, some species contained lysine and not diaminopimelic acid as the major dibasic amino acid (e.g., F. asaccharolyticus). It is concluded that peptidoglycans of gram-negative organisms constitute a somewhat more heterogeneous group than hitherto assumed.

Alanine↗