Liver microsomes contain multiple forms of inositol 1,4,5-trisphosphate binding proteins

Identification by nitrocellulose blot overlay

Research output: Contribution to journalArticle

Abstract

A group of proteins binding to inositol 1,4,5-trisphosphate (IP3) has been identified in rat liver microsomes by a nitrocellulose blot-overlay technique. Proteins were resolved by SDS-PAGE, blotted on nitrocellulose and incubated with [32P]IP3 followed by autoradiography. Approximately eight IP3-binding polypeptides ranging Mr 23-50 kDA were present exclusively in microsomes; these were absent from plasma membrane and mitochondrial fractions. Binding of [32P]IP3 to these proteins was displaceable to a great extent by 5 μM unlabeled IP3 but not by 10 μM IP1, IP2, IP4, ATP, or GTPγS. These results suggest that liver microsomes contain multiple forms of IP3-binding proteins that can be detected by this new method.

Original languageEnglish
Pages (from-to)79-83
Number of pages5
JournalJournal of Pharmacological and Toxicological Methods
Volume27
Issue number2
DOIs
StatePublished - 1992

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Inositol 1,4,5-Trisphosphate
Collodion
Liver Microsomes
Liver
Carrier Proteins
Cell membranes
Microsomes
Autoradiography
Protein Binding
Rats
Polyacrylamide Gel Electrophoresis
Proteins
Adenosine Triphosphate
Cell Membrane
Peptides

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Toxicology

Cite this

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title = "Liver microsomes contain multiple forms of inositol 1,4,5-trisphosphate binding proteins: Identification by nitrocellulose blot overlay",
abstract = "A group of proteins binding to inositol 1,4,5-trisphosphate (IP3) has been identified in rat liver microsomes by a nitrocellulose blot-overlay technique. Proteins were resolved by SDS-PAGE, blotted on nitrocellulose and incubated with [32P]IP3 followed by autoradiography. Approximately eight IP3-binding polypeptides ranging Mr 23-50 kDA were present exclusively in microsomes; these were absent from plasma membrane and mitochondrial fractions. Binding of [32P]IP3 to these proteins was displaceable to a great extent by 5 μM unlabeled IP3 but not by 10 μM IP1, IP2, IP4, ATP, or GTPγS. These results suggest that liver microsomes contain multiple forms of IP3-binding proteins that can be detected by this new method.",
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T1 - Liver microsomes contain multiple forms of inositol 1,4,5-trisphosphate binding proteins

T2 - Identification by nitrocellulose blot overlay

AU - Ali, Nawab

AU - Agrawal, Devendra K.

PY - 1992

Y1 - 1992

N2 - A group of proteins binding to inositol 1,4,5-trisphosphate (IP3) has been identified in rat liver microsomes by a nitrocellulose blot-overlay technique. Proteins were resolved by SDS-PAGE, blotted on nitrocellulose and incubated with [32P]IP3 followed by autoradiography. Approximately eight IP3-binding polypeptides ranging Mr 23-50 kDA were present exclusively in microsomes; these were absent from plasma membrane and mitochondrial fractions. Binding of [32P]IP3 to these proteins was displaceable to a great extent by 5 μM unlabeled IP3 but not by 10 μM IP1, IP2, IP4, ATP, or GTPγS. These results suggest that liver microsomes contain multiple forms of IP3-binding proteins that can be detected by this new method.

AB - A group of proteins binding to inositol 1,4,5-trisphosphate (IP3) has been identified in rat liver microsomes by a nitrocellulose blot-overlay technique. Proteins were resolved by SDS-PAGE, blotted on nitrocellulose and incubated with [32P]IP3 followed by autoradiography. Approximately eight IP3-binding polypeptides ranging Mr 23-50 kDA were present exclusively in microsomes; these were absent from plasma membrane and mitochondrial fractions. Binding of [32P]IP3 to these proteins was displaceable to a great extent by 5 μM unlabeled IP3 but not by 10 μM IP1, IP2, IP4, ATP, or GTPγS. These results suggest that liver microsomes contain multiple forms of IP3-binding proteins that can be detected by this new method.

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