A Putative Cytosolic Saxitoxin Binding Protein in Frog Myocardium

Approximately 20% of the protein in sarcolemma vesicles prepared from the hearts of the North American frog Rana pipiens pipiens is solubilized by a 96 hour dialysis of the vesicles against a low ionic strength solution of ethylene-diaminetetraacetic acid (EDTA) at pH 8.5. All of the polypeptides of the vesicle proteins which are resolvable by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 7.5% acrylamide disc gels are found in the aqueous supernatant (F1 fraction) when the dialysate is centrifuged at 100,000 xg for 1.5 hours. The yield of the individual proteins in F1 as revealed by the stained polypeptide bands on the gels varies. The total enzymatic activity of alkaline phosphatase and (Na('+) + K('+))-ATPase (ouabain sensitive) measured in the vesicles is retained throughout the EDTA treatment. 15% and 7% of their vesicular activities are found in F1 respectively. Total binding of Saxitoxin (STX), a Na('+)-channel specific neurotoxin, is only 25% retained, 6% in F1. Total 5'-nucleotidase activity is only 5% retained, less than 1% in F1. Sugar containing proteins detectable by periodic acid-Schiff base (PAS) staining of non-detergent (buffer) gels are found in F1. Proteins in F1 are associated with an appreciable amount of phospholipid. The protein-phospholipid complexes range in size from about 50,000 to several million daltons when measured by non-detergent gel electrophoresis.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biophysics

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Doyle, Donald David

• Biophysics, General

Language dc:language

eng

Identifier

(UMI)AAI8129572

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/67597