Labeling and isolation of plasma membranes from corn leaf protoplasts.

Plant physiology ·Vol. 65 ·No. 6 ·1980-06-00 ·Pages 1053-7

Perlin DS, Spanswick RM

Abstract

A plasma membrane-enriched fraction has been isolated from corn leaf mesophyll protoplasts and its identity confirmed with the aid of an external label, diazotized [(125)I]iodosulfanilic acid. Gentle cell disruption enabled internal organelles to be maintained intact and thus facilitated separation from the plasma membrane. The plasma membrane-enriched fraction was devoid of chloroplast or mitochondrial markers, whereas markers for the endoplasmic reticulum and golgi indicated minimal contamination. The highly enriched plasma membrane fraction contained a Mg(2+)-dependent, K(+)-stimulated ATPase with a pH optimum near neutrality. The position of the membranes on sucrose density gradients indicates that the plasma membranes have characteristics similar to other plasma membrane fractions.

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Perlin D S

Section of Botany, Genetics and Development, Division of Biological Sciences, Plant Science Building, Cornell University, Ithaca, New York 14853.

Spanswick R M

References (15)

15 references, click to expand

A FLUORESCENT LABEL FOR THE OUTER COMPONENTS OF THE PLASMA MEMBRANE.
Biochim Biophys Acta. 1964 Sep 25;88:390-9 PMID: 14253225
Cytoplasm-specific Effects of Helminthosporium maydis Race T Toxin on Survival of Corn Mesophyll Protoplasts.
Plant Physiol. 1978 Mar;61(3):420-4 PMID: 16660306
A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples.
Anal Biochem. 1978 Jun 15;87(1):206-10 PMID: 98070
The isolation of plasma membrane from protoplasts of soybean suspension cultures.
J Cell Sci. 1977 Apr;24:295-310 PMID: 561089
Diazontized (125I) diiodosulafanilic acid as a label for cell surface membranes. Studies on erythrocytes.
Biochim Biophys Acta. 1979 May 3;553(1):54-65 PMID: 454587
Purification of an ion-stimulated adenosine triphosphatase from plant roots: association with plasma membranes.
Proc Natl Acad Sci U S A. 1972 Nov;69(11):3307-11 PMID: 16592027
Evidence for the presence of the toxin-binding protein on the plasma membrane of sugarcane cells.
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1413-7 PMID: 4208550
ISOLATION OF beta-GLUCAN SYNTHETASE PARTICLES FROM PLANT CELLS AND IDENTIFICATION WITH GOLGI MEMBRANES.
Proc Natl Acad Sci U S A. 1969 Oct;64(2):605-12 PMID: 16591795
Adenosine triphosphatase from soybean callus and root cells.
Plant Physiol. 1977 Feb;59(2):264-7 PMID: 16659830
Purification of enzymatically isolated mesophyll protoplasts from c(3), c(4), and crassulacean Acid metabolism plants using an aqueous dextran-polyethylene glycol two-phase system.
Plant Physiol. 1973 Nov;52(5):484-90 PMID: 16658589
Differential Ion Stimulation of Plasmalemma Adenosine Triphosphatase from Leaf Epidermis and Mesophyll of Nicotiana rustica L.
Plant Physiol. 1979 May;63(5):936-9 PMID: 16660840
beta-Glucan Synthetases of Plasma Membrane and Golgi Apparatus from Onion Stem.
Plant Physiol. 1974 Sep;54(3):333-40 PMID: 16658884
Purification of a plasma membrane-bound adenosine triphosphatase from plant roots.
Methods Enzymol. 1974;32:392-406 PMID: 4155469
Cation-stimulated Adenosine Triphosphatase Activity and Cation Transport in Corn Roots.
Plant Physiol. 1976 Sep;58(3):331-5 PMID: 16659673
Isolation of plasma membranes from corn roots by sucrose density gradient centrifugation: an anomalous effect of ficoll.
Plant Physiol. 1976 Jan;57(1):105-14 PMID: 16659414

Article Info

Journal

Plant physiology

Abbr.

Plant Physiol

ISSN

0032-0889

DOI

10.1104/pp.65.6.1053

Published

1980-06-00

Pages

1053-7

Language

English

Region

United States

NLM ID

0401224

PMCID

PMC440479

External Links

PubMed source DOI: 10.1104/pp.65.6.1053 PMC Full Text View journal details

Analysis Services

Labeling and isolation of plasma membranes from corn leaf protoplasts.

Contact

WeChat Official Account