Properties of 3H-cimetidine binding in rat brain membrane fractions

The secretory vesicle cysteine string proteins (CSPs) are members of the DnaJ family of chaperones, and function at late stages of Ca2+-regulated exocytosis by an unknown mechanism. To determine novel binding partners of CSPs, we employed a pull-down strategy from purified rat brain membrane or cytosolic proteins using recombinant hexahistidine-tagged (His6-)CSP. Western blotting of the CSP-binding proteins identified synaptotagmin I to be a putative binding partner. Furthermore, pull-down assays using cAMP-dependent protein kinase (PKA)-phosphorylated CSP recovered significantly less synaptotagmin. Complexes containing CSP and synaptotagmin were immunoprecipitated from rat brain membranes, further suggesting that these proteins interact in vivo. Binding assays in vitro using recombinant proteins confirmed a direct interaction between the two proteins and demonstrated that the PKA-phosphorylated form of CSP binds synaptotagmin with approximately an order of magnitude lower affinity than the non-phosphorylated form. Genetic studies have implicated each of these proteins in the Ca2+-dependency of exocytosis and, since CSP does not bind Ca2+, this novel interaction might explain the Ca2+-dependent actions of CSP.

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Comparison of rat brain membrane antigens by complement mediated release of neurotransmitters from brain fractions using antisera against S-100 and THY-1

Brain Research ◽

10.1016/0006-8993(79)90131-8 ◽

1979 ◽

Vol 168 (1) ◽

pp. 119-132 ◽

Cited By ~ 10

Author(s):

K.G. Haglid ◽

A.N. Barclay ◽

A. Hamberger ◽

B. Hedquist ◽

B. Nystro¨m ◽

...

Keyword(s):

Rat Brain ◽

Brain Membrane ◽

S 100 ◽

Membrane Antigens

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Identification of a domain in cytochrome C that displaces [3H]TCP binding from rat brain membrane receptors: Synthesis of β-neuroprotectin

International Journal of Biochemistry ◽

10.1016/0020-711x(90)90135-p ◽

1990 ◽

Vol 22 (4) ◽

pp. 335-339 ◽

Cited By ~ 16

Author(s):

J.Blanche O'Neill ◽

H. Jaffe ◽

P.L. Hallberg ◽

J.M. Buzy ◽

T. Kingan ◽

...

Keyword(s):

Cytochrome C ◽

Rat Brain ◽

Membrane Receptors ◽

Brain Membrane

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Characterization of Phorbol Ester Binding to Membrane-Inserted Protein Kinase C in Rat Brain Membrane

Psychiatry and Clinical Neurosciences ◽

10.1111/j.1440-1819.1990.tb00457.x ◽

1990 ◽

Vol 44 (1) ◽

pp. 141-142

Author(s):

Ken'ichi Osada ◽

Mikio Asakura ◽

Makiko Shibata ◽

Tohru Tsukamoto ◽

Jun Imafuku ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Rat Brain ◽

Phorbol Ester ◽

Brain Membrane ◽

Kinase C

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Influence of oxygenated fuel additives and their metabolites on the binding of a convulsant ligand of the γ-aminobutyric acidA (GABAA) receptor in rat brain membrane preparations

Toxicology Letters ◽

10.1016/s0378-4274(02)00020-6 ◽

2002 ◽

Vol 129 (3) ◽

pp. 219-226 ◽

Cited By ~ 6

Author(s):

Joseph V Martin ◽

Nesli M Bilgin ◽

M.Michael Iba

Keyword(s):

Rat Brain ◽

Gabaa Receptor ◽

Fuel Additives ◽

Brain Membrane ◽

Oxygenated Fuel

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Comparative Stereostructure-Activity Studies on GABAA and GABAB Receptor Sites and GABA Uptake Using Rat Brain Membrane Preparations

Journal of Neurochemistry ◽

10.1111/j.1471-4159.1986.tb00695.x ◽

2006 ◽

Vol 47 (3) ◽

pp. 898-903 ◽

Cited By ~ 68

Author(s):

Erik Falch ◽

Anette Hedegaard ◽

Lone Nielsen ◽

Birger R. Jensen ◽

Hans Hjeds ◽

...

Keyword(s):

Rat Brain ◽

Gabab Receptor ◽

Gaba Uptake ◽

Brain Membrane ◽

Receptor Sites

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Imidazo[1,2-b]pyridazines. XII. Syntheses and Central Nervous System Activities of Some Substituted Imidazo[1,2-b]pyridazines and Related Imidazo[1,2-a]pyridines, Imidazo[1,2-a]pyrimidines and Imidazo[1,2-a]pyrazines

Australian Journal of Chemistry ◽

10.1071/ch9920877 ◽

1992 ◽

Vol 45 (5) ◽

pp. 877 ◽

Cited By ~ 15

Author(s):

GB Barlin ◽

LP Davies ◽

SJ Ireland ◽

MML Ngu ◽

JK Zhang

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Rat Brain ◽

Alkyl Group ◽

Significant Loss ◽

Aryl Group ◽

Brain Membrane ◽

Ic50 Values

Syntheses are reported for some 6-chloro(alkoxy,alkylthio and pheny1thio)-3-benzamidomethyl-(acetamidomethyl and methoxy)-2-arylimidazo[1,2-apyridines and some corresponding imidazo- [1,2- b]pyridazines, imidazo[1,2-a[pyrimidines and imidazo[l,2-a]pyrazines. IC50 values (or percentage displacement) are reported and discussed for the displacement of [3H]diazepam from rat brain membrane by each of these compounds. The imidazo[l,2-a[pyridines were generally slightly less potent than the imidazo[l,2- b]pyridazines but considerably more potent than the corresponding imidazo[1,2-a]pyrimidines or imidazo[1,2- a[pyrazines. Substitution of a 2-aryl group by a 2-alkyl group in imidazo[l,2- b]pyridazines led to a significant loss of activity.

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