Partial purification and characterization of a membrane-derived factor regulating neurotransmitter phenotypic expression. Academic Article uri icon

Overview

abstract

  • Cell membrane contact induces the de novo expression of choline O-acetyltransferase (CAT; acetyl-CoA: choline O-acetyltransferase, EC 2.3.1.6) activity in cultures of virtually pure neonatal rat dissociated sympathetic neurons. To identify molecular mechanisms underlying membrane-associated CAT induction, the responsible membrane component was characterized and partially purified. Substantial CAT-inducing activity was found in membranes from adult rat spinal cord and sensory and sympathetic ganglia. Whole brain membranes demonstrated significantly less activity. CAT induction in sympathetic neurons in response to spinal cord membranes was linear with respect to time, after an initial 6-hr lag. It was also linear with respect to concentrations of spinal cord protein from 2 to 100 micrograms per ml. CAT-inducing activity was extracted from spinal cord membranes by incubation with 100 mM NaCl and was purified approximately 5000-fold by DEAE ion-exchange and gel filtration chromatography. The active factor appears to be an extrinsic protein with an apparent molecular mass of 27 kDa. It is inactivated by trypsin and chymotrypsin but is moderately thermostable, retaining activity at 60 degrees C but not at 90 degrees C.

publication date

  • February 1, 1989

Research

keywords

  • Biological Factors
  • Choline O-Acetyltransferase
  • Ganglia, Sympathetic
  • Neurons
  • Neurotransmitter Agents

Identity

PubMed Central ID

  • PMC286625

Scopus Document Identifier

  • 0024502504

Digital Object Identifier (DOI)

  • 10.1073/pnas.86.3.1080

PubMed ID

  • 2563590

Additional Document Info

volume

  • 86

issue

  • 3