Isolation and partial characterization of human platelet vinculin
نویسندگان
چکیده
A 130,000 Mr protein was isolated from human platelets by sequential DEAE-Sephacel and Sepharose Cl-4B chromatography. Low shear viscometric measurements showed that the enriched protein after DEAE-Sephacel chromatography inhibited actin polymerization. This effect was somewhat greater in the presence of EGTA than in the presence of calcium. Further purification by Sepharose Cl-4B chromatography resulted in a complete loss of this inhibitory effect. Studies with fluorescent actin detected no nucleation or "+" end capping activity in either the DEAE-Sephacel- or Sepharose Cl-4B-purified vinculin. Antibodies raised in mice against the 130,000-mol-wt protein were shown to cross-react with chicken gizzard vinculin and a similar molecular weight protein was detected in WI38 cells and, Madin-Darby canine kidney cells. Lysis experiments with the Madin-Darby canine kidney cells indicated that most of the vinculin was soluble in Triton X-100, although some was found associated with the insoluble cytoskeletal residue. By immunofluorescence, vinculin in WI38 cells was localized to adhesion plaques as described by others. Discrete localization in platelets was also detected and appeared to depend on their state of adhesion and spreading. The results of these experiments suggest that human platelets contain a protein similar to vinculin. It is not clear if platelet vinculin is associated with structures analogous to adhesion plaques found in other cell types. The data indicate that the previously reported effects of nonmuscle vinculins on actin polymerization may be due to a contaminant or contaminants.
منابع مشابه
Isolation and characterization of Brachyspira species based on biochemical scheme and 16S rDNA partial sequencing
Avian intestinal spirochetosis (AIS) is a disease of birdscharacterized by a marked colonization of the cecum and/or rectumwith anaerobic intestinal bacteria of the genus Brachyspira. Thepurpose of the study was to determine the occurrence of Brachyspiraspp from avian sources in Iran and to characterize selected isolates bybiochemical and molecular methods. Out of 165 fecal samples obtained fro...
متن کاملThe Isolation and Characterization a Human Diploid Cell Strain and Its Use in Production of Measles Vaccine
متن کامل
Isolation and characterization of Phi class glutathione transferase partial gene from Iranian barley
Glutathione transferases are multifunctional proteins involved in several diverse intracellular events such as primary and secondary metabolisms, signaling and stress metabolism. These enzymes have been subdivided into eight classes in plants. The Phi class, being plant specific, is the most represented. In the present study, based on the sequences available at GenBank, different primers were d...
متن کاملIsolation and molecular characterization of partial FSH and LH receptor genes in Arabian camels (Camelus dromedarius)
Very little is known about LHR and FSHR genes of domestic dromedary camels. The main objective of this study was to determine and analyze partial genomic regions of FSHR and LHR genes in dromedary camels for the first time. To this end, a total of 50 DNA samples belonging to dromedary camels raised in Iran were sent for sequencing (25 samples of each gene). We compared the nucleotide sequences ...
متن کاملIsolation and partial characterization of a new strain of Klebsiella pneumoniae capable of high 1,3 propanediol production from glycerol
Glycerol is a promising feedstock for microbial cultivation and production of 1,3 propanediol (1,3 PDO). Here we report a newly isolated bacterial strain BA11 from soil, capable of fermenting glycerol to 1,3 PDO, and has been identified to be a strain of Klebsiella pneumoniae. Strain BA11 was fast growing showing peak 1,3 PDO production in 6 h of cultivation with productivity of 1.2 g/...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- The Journal of Cell Biology
دوره 100 شماره
صفحات -
تاریخ انتشار 1985