Detection of low molecular mass GTP-binding proteins in chromaffin granules and other subcellular fractions of chromaffin cells.

Abstract:

:A homogenate of purified chromaffin cells was fractionated, after removal of the nuclear fraction, by sucrose density gradient ultracentrifugation. The presence and subcellular localization of low molecular mass GTP-binding proteins was explored by incubation of blots of proteins from different subcellular fractions with [alpha-32P]GTP in the presence of Mg2+. The fractions enriched in intact chromaffin granule markers, i.e. catecholamines, chromogranin A, chromogranin B and cytochrome b-561 were also enriched in labelled GTP-binding proteins. Two major labelled components of 23 and 29 kDa were rapidly detected by autoradiography. Traces of 26 and 27 kDa components were also present. These components were detectable in both plasma and granule membranes. In addition to these components, the cytosolic fraction contained another GTP-binding protein of about 20 kDa. Binding of [alpha-32P]GTP was specific and dependent on Mg2+. By analogy to the findings reported in non-mammalian systems, the observations described here suggest the involvement of low molecular mass GTP-binding proteins in the chromaffin cell secretory process.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Doucet JP,Fournier S,Parulekar M,Trifaró JM

doi

10.1016/0014-5793(89)81254-2

subject

Has Abstract

pub_date

1989-04-10 00:00:00

pages

127-31

issue

1

eissn

0014-5793

issn

1873-3468

pii

0014-5793(89)81254-2

journal_volume

247

pub_type

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