Gordon A. Plishker

Regulation of plasma membrane Ca2+ ATPases by lipids of the phosphatidylinositol cycle

When the erythrocyte plasma membrane Ca2+ pump is reconstituted into phosphatidylcholine liposomes, the inclusion of small amounts of phosphatidic acid or phosphatidylinositol 4,5-bisphosphate stimulates the enzymes activity. Other lipids of the phosphatidylinositol cycle (diacylglycerol, phosphatidylinositol) have little effect. The stimulatory effect of phosphatidylinositol 4,5-bisphosphate is greater than that of calmodulin; this lipid also stimulates the plasma membrane Ca2+ ATPase from rat brain.

Bio-effects of high magnetic fields: A study using a simple animal model

The desire to do clinical imaging and spectroscopy at magnetic field strengths greater than 2 Tesla (T) necessitates investigation of possible bioeffects at these high fields. A simple T-maze was utilized to evaluate the aversive effects of exposure to three levels of static magnetic field (0, 1.5, and 4 T). The right arm of the maze extended into the center of a 30-cm horizontal bore magnet, while the left arm extended into a mock magnet bore with the same dimensions. The self-shielded design of the magnet reduces the fringe field to zero within 1 m of the bore, placing the start box of the maze outside the 5-G line of the magnet. Each rat performed a total of ten trials at each level of magnetic field strength. A follow-up subset was run at 4 T with the maze reversed. At 0 T, the rats entered the magnet freely. No significant differences from the control were observed at 1.5 T. At 4 T, however, in 97% of the trials the rats would not enter the magnet. In the maze-reversed subset a majority of the rats turned toward the magnet, indicating that they had learned an aversive response from the previous trials at 4 T. However, in only 4 decisions out of 58 did the rats actually enter the magnet. Eighteen decisions to turn around were made at the edge of the magnet in a region of strong field gradients (up to 13 T/m) and a field strength up to 1.75 T.(ABSTRACT TRUNCATED AT 250 WORDS)

Purification and measurement of calpromotin, the cytoplasmic protein which activates calcium-dependent potassium transport

A simple procedure is described for the purification of calpromotin, a protein from the cytoplasm of red blood cells which is capable of activating calcium-dependent potassium transport. The purification steps involve a salt gradient elution from an anion exchange column (Whatman DE-52) followed by a potassium phosphate gradient elution from a column of hydroxyapatite (HA Ultrogel). These steps result in a 54% yield with a 161 fold purification. The calpromotin is estimated to be 99% pure as determined by densitometry of the protein profile on an SDS polyacrylamide gel. A competitive enzyme-linked immunosorbent assay (ELISA) using rabbit anti-human calpromotin antibodies, is described for measuring levels of calpromotin in the 5 to 100 ng range.

Phenothiazine inhibition of calmodulin stimulates calcium-dependent potassium efflux in human red blood cells

Elevation of red blood cell calcium increases the efflux of potassium. The active extrusion of calcium from the red cell is regulated by calmodulin. Phenothiazines bind to calmodulin in a calcium-dependent manner preventing the calmodulin from activating a wide variety of cellular processes. The present study shows that phenothiazines increase the efflux of potassium from red cells incubated with the calcium ionophore A23187. The dose dependent effect of trifluoperazine on potassium efflux correlates with its inhibition of Ca-ATPase activity. The phenothiazine effects are dependent upon ATP in that increases in potassium efflux are not observed in energy depleted cells. In calcium buffered ghosts no direct effect of calmodulin or an antibody to calmodulin can be shown. These data suggest that phenothiazines stimulate calcium-dependent potassium loss indirectly by a drug-induced blockage of the calmodulin-activated Ca-ATPase.

Calcium-dependent association of glutathione S-transferase with the human erythrocyte membrane

Elevations in intracellular calcium increase the adsorption of a cytoplasmic protein to human red blood cell membrane. This protein migrates on SDS polyacrylamide gels at 23,000 daltons and has been called band 8. The association of this protein with the membrane is increased in sickle cell anemia. This protein is extracted from the membrane with EGTA, a calcium chelator. Enzymatic and immunological studies identify band 8 as a glutathione S-transferase.

Inherited Membrane Disorders of Muscle

Myotonic (MyD) and Duchenne (DMD) muscular dystrophy are two of the more common and devastating inherited muscular dystrophies in man. In this chapter we will review MyD and DMD in terms of the pertinent clinical data and present a review of the recent electrophysiological, morphological, and biochemical findings in these two disorders.

Phosphoglyceride biosynthesis in bovine adrenal chromaffin cells

Cultured adrenal chromaffin cells, representing a virtually homogeneous population of neuronai elements, have been utilized to examine the final enzymes in the formation of phosphatidylcholine (PC) and phosphatidylethanolamine (PE), namely, choline phosphotransferase, ethanolaminephosphotransferase, and the N-methyltransferases in the sequential methylation of PE to PC. Each enzyme has been characterized extensively in terms of substrate requirements, pH optima, detergent and cation effects, and response to inhibitors revealing properties very similar to those in other neural preparations. The respective activities are stable for up to two weeks of adrenal chromaffin cell culture suggesting that this system is a suitable model for examining the relative roles and the regulation of each pathway in PC formation.