Measurement of proton-linked transport activities in pyranine loaded chloroplast inner envelope vesicles.

Abstract:

:A method has been devised for loading chloroplast inner envelope vesicles prepared from pea (Pisum sativum L. var Progress No. 9) or spinach (Spinacia oleracea L.) with 8-hydroxypyrene-1,3,6-trisulfonate (pyranine), a membrane impermeant, fluorescent pH indicator. Two known proton-linked transport activities of the inner envelope, glycolate/H(+) co-transport and phosphate/phosphoglycerate exchange have been shown to cause quenching of the internal pyranine fluorescence. This represents the first demonstration that these vesicles are sealed and competent for transport measurements. The technique, as it now stands, is essentially qualitative. It does, however, offer advantages over transport measurements with intact chloroplasts, for example compatibility with rapid mixing techniques and accessibility of the transport proteins to antibodies.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Howitz KT,McCarty RE

doi

10.1104/pp.86.4.999

subject

Has Abstract

pub_date

1988-04-01 00:00:00

pages

999-1001

issue

4

eissn

0032-0889

issn

1532-2548

journal_volume

86

pub_type

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