Document Detail


Activation of the human red cell calcium ATPase by calcium pretreatment.
MedLine Citation:
PMID:  8182735     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Some kinetic parameters of the human red cell Ca(2+)-ATPase were studied on calmodulin-free membrane fragments following preincubation at 37 degrees C. After 30 min treatment with EGTA (1 mM) plus dithioerythritol (1 mM), a Vmax of about 0.4 mumol Pi/mg x hr and a Ks of 0.3 microM Ca2+ were found. When Mg2+ (10 mM) or Ca2+ (10 microM) were also added during preincubation, Vmax, but not Ks was altered. Ca2+ was more effective than Mg2+, thus increasing Vmax to about 1.3 mumol P/mg x hr. The presence of both Ca2+ and Mg2+ during pretreatment decreased Ks to 0.15 microM, while having no apparent effect on Vmax. Conversely, addition of ATP (2 mM) with either Ca2+ or Ca2+ plus Mg2+ increased Vmax without affecting Ks. Preincubation with Ca2+ for periods longer than 30 min further increased Vmax and reduced Ks to levels as low as found with calmodulin treatment. The Ca2+ activation was not prevented by adding proteinase inhibitors (iodoacetamide, 10 mM; leupeptin, 200 microM; pepstatin A, 100 microM; phenylmethanesulfonyl fluoride, 100 microM). The electrophoretic pattern of membranes preincubated with or without Mg2+, Ca2+ or Ca2+ plus Mg2+ did not differ significantly from each other. Moreover, immunodetection of Ca(2+)-ATPase by means of polyclonal antibodies revealed no mobility change after the various treatments. The above stimulation was not altered by neomycin (200 microM), washing with EGTA (5 mM) or by both incubating and washing with delipidized serum albumin (1 mg/ml), or omitting dithioerythritol from the preincubation medium.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
J Fermin; P J Romero
Related Documents :
8780185 - Effects of mg2+ on ca2+ waves and ca2+ transients of rat ventricular myocytes.
3136805 - Regulation of the microtubule-lysosome interaction: activation by mg2+ and inhibition b...
7441155 - The role of divalent cations in the metabolic response of mouse blastocysts to serum.
2912705 - The effects of high extracellular ca2+ and mg2+ concentrations on the levels of inosito...
2211025 - Single potassium channels in corneal epithelium.
1979145 - Mechanism of the cardiotoxic action of palytoxin.
Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of membrane biology     Volume:  137     ISSN:  0022-2631     ISO Abbreviation:  J. Membr. Biol.     Publication Date:  1994 Feb 
Date Detail:
Created Date:  1994-06-16     Completed Date:  1994-06-16     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0211301     Medline TA:  J Membr Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  271-7     Citation Subset:  IM    
Affiliation:
Instituto de Biología Experimental, Fac. Ciencias, Universidad Central de Venezuela, Caracas.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / pharmacology
Calcium / pharmacology*
Calcium-Transporting ATPases / blood*,  chemistry
Egtazic Acid / pharmacology
Enzyme Activation / drug effects
Erythrocyte Membrane / drug effects*,  enzymology*
Humans
Kinetics
Magnesium / pharmacology
Membrane Fluidity / drug effects
Neomycin / pharmacology
Protein Conformation / drug effects
Chemical
Reg. No./Substance:
1404-04-2/Neomycin; 56-65-5/Adenosine Triphosphate; 67-42-5/Egtazic Acid; 7439-95-4/Magnesium; 7440-70-2/Calcium; EC 3.6.1.8/Calcium-Transporting ATPases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Analysis of the distribution and phosphorylation state of ZO-1 in MDCK and nonepithelial cells.
Next Document:  Which nucleotides in the "-10" region are crucial to obtain a fully active MalT-dependent ...