A 75-kDa Polypeptide, Located Primarily at the Plasma Membrane of Carrot Cell-suspension Cultures, is Photoaffinity Labeled by the Calcium Channel Blocker LU 49888

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1990 Dec 15

PMID 11607139

Citations 8

Authors

P Thuleau

A Graziana

H Canut

R Ranjeva

Affiliations

Soon will be listed here.

Abstract

Calcium channel blockers of the phenylalkylamine family bind specifically to membranes and inhibit calcium uptake in carrot protoplast. LU 49888, an azido derivative of phenylalkylamine, behaves as its unmodified homolog in terms of affinity and specificity and therefore allows us to probe the receptor by photoaffinity labeling. Upon UV irradiation, a 75-kDa peptide was specifically labeled. Incubation of microsomes with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, a zwitterionic detergent, led to the solubilization of the LU 49888-binding protein. Electrophoretic analysis under denaturing conditions and gel filtration of the solubilized "receptor-ligand" complex show a 75-kDa peptide mainly located at the plasma membrane. Consequently the LU 49888-binding protein in plants differs significantly from its animal counterpart by its size and may be a primary target for external signal molecules.

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Solubilized proteins from carrot (Daucus carota L.) membranes bind calcium channel blockers and form calcium-permeable ion channels.

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References

Mikami A, Imoto K, Tanabe T, Niidome T, Mori Y, Takeshima H . Primary structure and functional expression of the cardiac dihydropyridine-sensitive calcium channel. Nature. 1989; 340(6230):230-3. DOI: 10.1038/340230a0. View

Andrejauskas E, Hertel R, Marme D . Specific binding of the calcium antagonist [3H]verapamil to membrane fractions from plants. J Biol Chem. 1985; 260(9):5411-4. View

Vaghy P, Striessnig J, Miwa K, Knaus H, Itagaki K, McKenna E . Identification of a novel 1,4-dihydropyridine- and phenylalkylamine-binding polypeptide in calcium channel preparations. J Biol Chem. 1987; 262(29):14337-42. View

Striessnig J, Knaus H, Grabner M, Moosburger K, Seitz W, LIETZ H . Photoaffinity labelling of the phenylalkylamine receptor of the skeletal muscle transverse-tubule calcium channel. FEBS Lett. 1987; 212(2):247-53. DOI: 10.1016/0014-5793(87)81354-6. View

Ferry D, Kampf K, Goll A, Glossmann H . Subunit composition of skeletal muscle transverse tubule calcium channels evaluated with the 1,4-dihydropyridine photoaffinity probe, [3H]azidopine. EMBO J. 1985; 4(8):1933-40. PMC: 554443. DOI: 10.1002/j.1460-2075.1985.tb03873.x. View

Harvey H, Venis M, Trewavas A . Partial purification of a protein from maize (Zea mays) coleoptile membranes binding the Ca2+-channel antagonist verapamil. Biochem J. 1989; 257(1):95-100. PMC: 1135542. DOI: 10.1042/bj2570095. View

Smith P, Krohn R, Hermanson G, Mallia A, Gartner F, Provenzano M . Measurement of protein using bicinchoninic acid. Anal Biochem. 1985; 150(1):76-85. DOI: 10.1016/0003-2697(85)90442-7. View

Striessnig J, Scheffauer F, Mitterdorfer J, Schirmer M, Glossmann H . Identification of the benzothiazepine-binding polypeptide of skeletal muscle calcium channels with (+)-cis-azidodiltiazem and anti-ligand antibodies. J Biol Chem. 1990; 265(1):363-70. View

Skinner M, Griswold M . Fluorographic detection of radioactivity in polyacrylamide gels with 2,5-diphenyloxazole in acetic acid and its comparison with existing procedures. Biochem J. 1983; 209(1):281-4. PMC: 1154087. DOI: 10.1042/bj2090281. View

10.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

11.

Conrad P, Hepler P . The effect of 1,4-dihydropyridines on the initiation and development of gametophore buds in the moss funaria. Plant Physiol. 1988; 86(3):684-7. PMC: 1054552. DOI: 10.1104/pp.86.3.684. View

12.

Sandelius A, Penel C, Auderset G, Brightman A, Millard M, Morre D . Isolation of highly purified fractions of plasma membrane and tonoplast from the same homogenate of soybean hypocotyls by free-flow electrophoresis. Plant Physiol. 1986; 81(1):177-85. PMC: 1075303. DOI: 10.1104/pp.81.1.177. View

13.

Jones A, Venis M . Photoaffinity labeling of indole-3-acetic acid-binding proteins in maize. Proc Natl Acad Sci U S A. 1989; 86(16):6153-6. PMC: 297795. DOI: 10.1073/pnas.86.16.6153. View

14.

Thuleau P, Graziana A, Rossignol M, Kauss H, Auriol P, Ranjeva R . Binding of the phytotoxin zinniol stimulates the entry of calcium into plant protoplasts. Proc Natl Acad Sci U S A. 1988; 85(16):5932-5. PMC: 281879. DOI: 10.1073/pnas.85.16.5932. View

15.

Hicks G, Rayle D, Jones A, Lomax T . Specific photoaffinity labeling of two plasma membrane polypeptides with an azido auxin. Proc Natl Acad Sci U S A. 1989; 86:4948-52. PMC: 297533. DOI: 10.1073/pnas.86.13.4948. View

16.

Schneider T, Hofmann F . The bovine cardiac receptor for calcium channel blockers is a 195-kDa protein. Eur J Biochem. 1988; 174(2):369-75. DOI: 10.1111/j.1432-1033.1988.tb14107.x. View

17.

Galizzi J, Borsotto M, Barhanin J, Fosset M, Lazdunski M . Characterization and photoaffinity labeling of receptor sites for the Ca2+ channel inhibitors d-cis-diltiazem, (+/-)-bepridil, desmethoxyverapamil, and (+)-PN 200-110 in skeletal muscle transverse tubule membranes. J Biol Chem. 1986; 261(3):1393-7. View

18.

Campbell K, Leung A, Sharp A . The biochemistry and molecular biology of the dihydropyridine-sensitive calcium channel. Trends Neurosci. 1988; 11(10):425-30. DOI: 10.1016/0166-2236(88)90193-2. View