A Cerebellar Purkinje Cell Marker P400 Protein is an Inositol 1,4,5-trisphosphate (InsP3) Receptor Protein. Purification and Characterization of InsP3 Receptor Complex

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1990 Jan 1

PMID 2153079

Citations 60

Authors

N Maeda

M Niinobe

K Mikoshiba

Affiliations

Soon will be listed here.

Abstract

P400 protein is a 250 kd glycoprotein, characteristic of the cerebellum, which is accumulated at the endoplasmic reticulum, at the plasma membrane and at the post-synaptic density of Purkinje cells. In this study, we purified inositol 1,4,5-trisphosphate (InsP3) receptor from mouse cerebellum and examined the possibility that P400 protein is identical with cerebellar InsP3 receptor protein. InsP3 receptor was solubilized with Triton X-100 from a post-nuclear fraction of ddY mouse cerebellum and was purified with high yield by sequential column chromatography on DE52, heparin-agarose, lentil lectin-Sepharose and hydroxylapatite. In these chromatographies, P400 protein co-migrated completely with the InsP3 binding activity. The purified receptor is a 250 kd protein with a Bmax of 2.1 pmol/microgram and a KD of 83 nM. It reacted with three different monoclonal antibodies against P400 protein, indicating that P400 protein is the same substance as the InsP3 receptor (P400/InsP3 receptor protein). Electron microscopy of the purified receptor showed a square shape with sides approximately 25 nm long. Binding assays of the cerebella of Purkinje cell-degeneration (pcd) mice with [3H]InsP3 demonstrated that the InsP3 binding sites in the cerebellum are distributed exclusively on the Purkinje cells. Immunohistochemical analysis indicated that P400/InsP3 receptor is present at the dendrites, cell bodies, axons and synaptic boutons of the Purkinje cells.

Citing Articles

Conformational motions and ligand-binding underlying gating and regulation in IPR channel.

Fan G, Baker M, Terry L, Arige V, Chen M, Seryshev A Nat Commun. 2022; 13(1):6942.

PMID: 36376291 PMC: 9663519. DOI: 10.1038/s41467-022-34574-1.

Neuronal microRNAs safeguard ER Ca homeostasis and attenuate the unfolded protein response upon stress.

Paschou M, Papazafiri P, Charalampous C, Zachariadis M, Dedos S, Doxakis E Cell Mol Life Sci. 2022; 79(7):373.

PMID: 35727337 PMC: 11073139. DOI: 10.1007/s00018-022-04398-9.

Lipid Dyshomeostasis and Inherited Cerebellar Ataxia.

Zhao J, Zhang H, Fan X, Yu X, Huai J Mol Neurobiol. 2022; 59(6):3800-3828.

PMID: 35420383 PMC: 9148275. DOI: 10.1007/s12035-022-02826-2.

Emerging Roles of Calcium Signaling in the Development of Non-Alcoholic Fatty Liver Disease.

Chen C, Hsu L, Chen K, Chiu K, Chen C, Huang K Int J Mol Sci. 2022; 23(1).

PMID: 35008682 PMC: 8745268. DOI: 10.3390/ijms23010256.

Inositol 1,4,5-Trisphosphate Receptors in Human Disease: A Comprehensive Update.

Gambardella J, Lombardi A, Morelli M, Ferrara J, Santulli G J Clin Med. 2020; 9(4).

PMID: 32290556 PMC: 7231134. DOI: 10.3390/jcm9041096.

References

Yano K, Higashida H, Inoue R, Nozawa Y . Bradykinin-induced rapid breakdown of phosphatidylinositol 4,5-bisphosphate in neuroblastoma X glioma hybrid NG108-15 cells. J Biol Chem. 1984; 259(16):10201-7. View

Berridge M, Irvine R . Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984; 312(5992):315-21. DOI: 10.1038/312315a0. View

Mikoshiba K, Okano H, Tsukada Y . P400 protein characteristic to Purkinje cells and related proteins in cerebella from neuropathological mutant mice: autoradiographic study by 14C-leucine and phosphorylation. Dev Neurosci. 1985; 7(3):179-87. DOI: 10.1159/000112286. View

Inui M, Saito A, Fleischer S . Purification of the ryanodine receptor and identity with feet structures of junctional terminal cisternae of sarcoplasmic reticulum from fast skeletal muscle. J Biol Chem. 1987; 262(4):1740-7. View

Kano M, Kato M . Quisqualate receptors are specifically involved in cerebellar synaptic plasticity. Nature. 1987; 325(6101):276-9. DOI: 10.1038/325276a0. View

Worley P, Baraban J, Colvin J, Snyder S . Inositol trisphosphate receptor localization in brain: variable stoichiometry with protein kinase C. Nature. 1987; 325(7000):159-61. DOI: 10.1038/325159a0. View

Kuno M, Gardner P . Ion channels activated by inositol 1,4,5-trisphosphate in plasma membrane of human T-lymphocytes. Nature. 1987; 326(6110):301-4. DOI: 10.1038/326301a0. View

Berridge M . Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem. 1987; 56:159-93. DOI: 10.1146/annurev.bi.56.070187.001111. View

Hirano A, DEMBITZER H . The fine structure of staggerer cerebellum. J Neuropathol Exp Neurol. 1975; 34(1):1-11. DOI: 10.1097/00005072-197501000-00001. View

10.

Mallet J, Huchet M, Pougeois R, Changeux J . Anatomical, physiological and biochemical studies on the cerebellum from mutant mice. III. Protein differences associated with the weaver, staggerer and nervous mutations. Brain Res. 1976; 103(2):291-312. DOI: 10.1016/0006-8993(76)90800-3. View

11.

Mikoshiba K, Changeux J . Morphological and biochemical studies on isolated molecular and granular layers from bovine cerebellum. Brain Res. 1978; 142(3):487-504. DOI: 10.1016/0006-8993(78)90911-3. View

12.

Mikoshiba K, Huchet M, Changeux J . Biochemical and immunological studies on the P400 protein, a protein characteristic of the Purkinje cell from mouse and rat cerebellum. Dev Neurosci. 1979; 2(6):254-75. DOI: 10.1159/000112489. View

13.

Kornfeld K, Reitman M, KORNFELD R . The carbohydrate-binding specificity of pea and lentil lectins. Fucose is an important determinant. J Biol Chem. 1981; 256(13):6633-40. View

14.

Ito M, Sakurai M, Tongroach P . Climbing fibre induced depression of both mossy fibre responsiveness and glutamate sensitivity of cerebellar Purkinje cells. J Physiol. 1982; 324:113-34. PMC: 1250696. DOI: 10.1113/jphysiol.1982.sp014103. View

15.

Ito M, Kano M . Long-lasting depression of parallel fiber-Purkinje cell transmission induced by conjunctive stimulation of parallel fibers and climbing fibers in the cerebellar cortex. Neurosci Lett. 1982; 33(3):253-8. DOI: 10.1016/0304-3940(82)90380-9. View

16.

Crepel F, Dupont J, Gardette R . Selective absence of calcium spikes in Purkinje cells of staggerer mutant mice in cerebellar slices maintained in vitro. J Physiol. 1984; 346:111-25. PMC: 1199487. DOI: 10.1113/jphysiol.1984.sp015010. View

17.

Worley P, Baraban J, Supattapone S, Wilson V, Snyder S . Characterization of inositol trisphosphate receptor binding in brain. Regulation by pH and calcium. J Biol Chem. 1987; 262(25):12132-6. View

18.

Shah J, Cohen R, Pant H . Inositol trisphosphate-induced calcium release in brain microsomes. Brain Res. 1987; 419(1-2):1-6. DOI: 10.1016/0006-8993(87)90562-2. View

19.

Supattapone S, Worley P, Baraban J, Snyder S . Solubilization, purification, and characterization of an inositol trisphosphate receptor. J Biol Chem. 1988; 263(3):1530-4. View

20.

Mattson M . Neurotransmitters in the regulation of neuronal cytoarchitecture. Brain Res. 1988; 472(2):179-212. DOI: 10.1016/0165-0173(88)90020-3. View