scholarly journals Isolation and partial characterization of pancreatic polypeptide-like material in the brain of the blowfly Calliphora vomitoria

1981 ◽  
Vol 197 (3) ◽  
pp. 767-770 ◽  
Author(s):  
H Duve ◽  
A Thorpe ◽  
R Neville ◽  
N R Lazarus
Keyword(s):  
Cyclic Amp ◽  
Pancreatic Polypeptide ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Structural Similarity ◽  
Cross Reactivity ◽  
Polyacrylamide Gel ◽  
Bovine Pancreatic Polypeptide ◽  
The Brain

Using 10(6) flies (5 kg of heads) a pancreatic polypeptide-like material has been partially purified from the blowfly Calliphora vomitoria. The isolated material was eluted on Sephadex G-50 similarly to bovine pancreatic polypeptide and had an RF on polyacrylamide-gel electrophoresis that was identical with that of the bovine hormone. The material diluted linearly and showed parallelism with bovine standards in a bovine pancreatic polypeptide immunoassay. In specificity controls the immunoreactivity was not abolished by trasylol and no cross-reactivity was discerned in assay for glucagon, proangiotensin and cyclic AMP. These data suggest that the pancreatic polypeptide material in the brain of the blowfly has close structural similarity to the mammalian hormone.

Purification And Characterization Of Calmodulin From Bovine Platelet

1981 ◽  
Author(s):  
J Kambayashi ◽  
M Sakon ◽  
G Kōsaki ◽  
K Sobue ◽  
S Kakiuchi
Keyword(s):  
Gel Electrophoresis ◽  
Blood Platelets ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Purification Method ◽  
Platelet Reaction ◽  
Platelet Pellet ◽  
The Brain

Calmodulin, which was originally reported as a Ca2+ dependent activator of phosphodiesterase, mediates a number of actions of Ca2+ as a second messenger for stimulus linked cellular responces in eukaryotic cells. In blood platelets, the availability of Ca2+is a prerequisite for steps of platelet reaction leading to the formation of hemostatic plug. The presence of this protein in platelets has been reported. In the present study, calmodulin was purified from bovine platelets and was extensively characterized.The identification of the protein was based on the activity to activate the brain calmodulin deficient phosphodiesterase. Calmodulin was purified 600 fold from bovine platelet pellet by a 3-step purification method consisted of trichloroacetic acid treatment, DEAE-cellulose column chromatography and phenothiazine affinity chromatography. The recovery rate was more than 70% and the purified protein was homogenous on polyacrylamide gel electrophoresis. The mobility of the purified material upon polyacrylamide gel electrophoresis was identical with that of brain calmodulin in the presence of either Ca2+or EGTA. In the quantitative analysis of the phosphodiesterase activation, the behaviour of the purified protein was also identical with brain calmodulin.From these strict characterization, it may be concluded that the purified protein is indistinguishable from brain calmodulin. The amount of the protein in bovine platelet was estimated to be 63 micrograms per wet gram, which is about one sixth of the amount in brain tissue.

Prothrombin Metz : Purification and Characterization of a Variant of Human Prothrombin

1979 ◽  
Author(s):  
M Ribieto ◽  
J Elion ◽  
D Labie ◽  
F Josso
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Xa ◽  
Polyacrylamide Gel ◽  
Cleavage Pattern ◽  
Purification And Characterization ◽  
Double Immunodiffusion ◽  
The Family

For the purification of the abnormal prothrombin (Pt Metz), advantage has been taken of the existence in the family of three siblings who, being double heterozygotes for Pt Metz and a hypoprothrombinemia, have no normal Pt. Purification procedures included barium citrate adsorption and chromatography on DEAE Sephadex as for normal Pt. As opposed to some other variants (Pt Barcelona and Madrid), Pt Metz elutes as a single symetrical peak. By SDS polyacrylamide gel electrophoresis, this material is homogeneous and appears to have the same molecular weight as normal Pt. Comigration of normal and abnormal Pt in the absence of SDS, shows a double band suggesting an abnormal charge for the variant. Pt Metz exhibits an identity reaction with the control by double immunodiffusion. Upon activation by factor Xa, Pt Metz can generate amydolytic activity on Bz-Phe-Val-Arg-pNa (S2160), but only a very low clotting activity. Clear abnormalities are observed in the cleavage pattern of Pt Metz when monitored by SDS gel electrophoresis. The main feature are the accumulation of prethrombin l (Pl) and the appearance of abnormal intermediates migrating faster than Pl.

Characterization of immunoreactive proteins of Setaria cervi isolated by preparative polyacrylamide gel electrophoresis

2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Priyanka Priyadarshi ◽  
Piyush Dravid ◽  
Inayat Hussain Sheikh ◽  
Sunita Saxena ◽  
Ashish Tandon ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Complex Mixtures ◽  
Setaria Cervi ◽  
Antigenic Proteins ◽  
Specific Antigens

AbstractFilarial parasites are complex mixtures of antigenic proteins and characterization of these antigenic molecules is essential to identify the diagnostically important filaria-specific antigens. In the present study, we have fractionated the somatic extracts from adults of

scholarly journals Phosphorylation of synaptic-membrane proteins from ox cerebral cortex in vitro. Preparation of fractions enriched in phosphorylated proteins by using extraction with detergents and urea, and gel filtration

1977 ◽  
Vol 163 (2) ◽  
pp. 369-378 ◽  
Author(s):  
P R Dunkley ◽  
H Holmes ◽  
R Rodnight
Keyword(s):  
Cerebral Cortex ◽  
Cyclic Amp ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Synaptic Membrane ◽  
Hydroxyl Groups ◽  
Phosphorylated Proteins

Synaptic-membrane fragments from ox cerebral cortex contain basal and cyclic AMP-stimulated protein kinase(s) that transfer 32P from [gamma-32P]ATP to hydroxyl groups of serine and threonine residues in membrane-protein substrates. In the present work, labelled membrane fragments were partitioned into soluble and insoluble fractions with Triton X-100, Nonidet P. 40, sodium deoxycholate and urea, and the distribution of 32P-labelled protein in the fractions was determined by polyacrylamide-gel electrophoresis and radioautography. A high percentage of phosphorylated protein sustrates remained insoluble, including those whose phosphorylation was most highly stimulated by cyclic AMP. Whole membrane fragments and samples prepared by detergent extraction were fractionated on Sepharose 6B columns in the presence of low concentrations of sodium dodecyl sulphate and pooled fractions were analysed by polyacrylamide-gel electrophoresis and radioautography. Phosphorylated proteins were fractionated on the basis of their molecular weight, but homogeneous protein was not obtained. The results are discussed in relation to the techniques used and the results obtained in other laboratories.

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