+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

High affinity binding of 9-tritiated methyl-7-bromoeudistomin D to the caffeine-binding site of skeletal muscle sarcoplasmic reticulum



High affinity binding of 9-tritiated methyl-7-bromoeudistomin D to the caffeine-binding site of skeletal muscle sarcoplasmic reticulum



Journal of Biological Chemistry 268(25): 18622-18625



3H-Labeled 9-methyl-7-bromoeudistomin D ((3H) MBED), the most powerful inducer of Ca-2+ release from sarcoplasmic reticulum (SR), was successfully prepared with a high specific activity of 10.2 Ci/mmol. (3H)MBED bound to terminal cisternae (TC) of skeletal muscle SR in a replaceable and saturable manner, indicating the existence of its specific binding site. Caffeine inhibited the (3H)MBED binding to the TC-SR membranes from skeletal muscle with an IC-50 value of 0.8 mM, in close agreement with a concentration that causes Ca-2+ release from SR. Scatchard analysis gave values of K-D = 40 nM and B-max = 10 pmol/mg protein. The K-D value was increased by caffeine, while that of B-max was not changed, indicating a competitive mode of inhibition. Adenosine 5'-(beta,gamma-methylene)triphosphate enhanced (3H)MBED binding, but ryanodine and Ca-2+ did not affect it. (3H)MBED binding to TC-SR membranes was inhibited by procaine, a representative blocker of Ca-2+-induced Ca-2+ release channels, whereas that was not changed by Mg-2+, suggesting that procaine but not Mg-2+ may exert its inhibitory effect on Ca-2+-induced Ca-2+ release by affecting the caffeine-binding sites. These results suggest that MBED shares the same binding site as that of caffeine in TC-SR. The (3H)MBED is the first radiolabeled ligand for caffeine-binding sites in Ca-2+ release channels and thus may provide an essential biochemical tool for elucidating this site.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 008770386

Download citation: RISBibTeXText


Related references

High affinity binding of 9-[3H]methyl-7-bromoeudistomin D to the caffeine-binding site of skeletal muscle sarcoplasmic reticulum. Journal of Biological Chemistry 268(25): 18622, 1993

The specific binding site of 9-[3H]methyl-7-bromoeudistomin D, a caffeine-like Ca2+ releaser, in liver microsomes in distinct from that in skeletal sarcoplasmic reticulum. Biological Chemistry Hoppe-Seyler 375(3): 183-187, 1994

The Specific Binding Site of 9-[ 3 H]Methyl-7-bromoeudistomin D, a Caffeine-Like Ca 2+ Releaser, in Liver Microsomes is Distinct from that in Skeletal Sarcoplasmic Reticulum. Biological Chemistry Hoppe-Seyler 375(3): 183-188, 1994

The specific binding site of 9-(3H) Methyl-7-bromoeudistomin D, a caffeine-like Ca-2+ releaser, in liver microsomes is distinct from that in skeletal sarcoplasmic reticulum. Biological Chemistry Hoppe Seyler 375(3): 183-187, 1994

Nd3+ and Co2+ binding to sarcoplasmic reticulum CaATPase. An estimation of the distance from the ATP binding site to the high-affinity calcium binding sites. Journal of Biological Chemistry 259(23): 14651, 1984

9-Methyl-7-bromoeudistomin D, a potent inducer of calcium release from sarcoplasmic reticulum of skeletal muscle. Experientia 45(8): 782-783, 1989

Molecular cloning of the high affinity calcium-binding protein (calreticulin) of skeletal muscle sarcoplasmic reticulum. Journal of Biological Chemistry 264(36): 21522, 1989

Low-affinity Ca(2+)-binding sites versus Zn(2+)-binding sites in histidine-rich Ca(2+)-binding protein of skeletal muscle sarcoplasmic reticulum. Biochemical and Biophysical Research Communications 186(2): 659-667, 1992

Assembly of the sarcoplasmic reticulum. Biosynthesis of the high affinity calcium binding protein in rat skeletal muscle cell cultures. Journal of Biological Chemistry 255(4): 1327-1334, 1980

Approach to clarification of caffeine binding sites in sarcoplasmic reticulum properties of tritiated mbed binding. Japanese Journal of Pharmacology 52(Suppl. 1): 35P, 1990

Neodymium and cobalt binding to sarcoplasmic reticulum calcium atpase an estimation of the distance from the atp binding site to the high affinity calcium binding sites. Journal of Biological Chemistry 259(23): 14651-14656, 1984

Effect of temperature on tritiated ryanodine binding to sarcoplasmic reticulum from bullfrog skeletal muscle. Nishizuka, Y , M Endo And C Tanaka (Ed ) Advances in Second Messenger And Phosphoprotein Research, Vol 24 The Biology And Medicine Of Signal Transduction; 7th International Conference on Cyclic Nucleotides, Calcium And Protein Phosphorylation, Kobe, Japan, October 8-13, 1989 Xxxiii+750p Raven Press: New York, New York, Usa Illus 605, 1990

Tritiated ryanodine binding to chicken fast twitch skeletal muscle sarcoplasmic reticulum. Japanese Journal of Pharmacology 52(Suppl. 1): 85P, 1990

Tritiated ryanodine binding to avian fast twitch skeletal muscle sarcoplasmic reticulum sr membranes. FASEB Journal 3(4): A1198, 1989

Effect of temperature on tritiated ryanodine binding to sarcoplasmic reticulum isolated from bullfrog skeletal muscle. Japanese Journal of Pharmacology 49(Suppl.): 93P, 1989