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. 1977 Nov;60(5):767–770. doi: 10.1104/pp.60.5.767

Isolation of Envelope Membranes from Bundle Sheath Chloroplasts of Maize

Raymond P Poincelot a,1
PMCID: PMC542711  PMID: 16660181

Abstract

Bundle sheath strands were isolated from maize (Zea mays L.) leaves treated with preparations of cellulase, hemicellulase, and pectinase. A three-phase discontinuous gradient yielded two fractions of envelope membranes from bundle sheath chloroplasts. Buoyant densities were 1.06 and 1.09 g cm−3. The lighter fraction contained membrane vesicles under light microscopy, but centrifugation produced a pellet that was too small and unstable for purposes of electron microscopy. The heavier fraction contained single and double membrane vesicles and was studied further. Enzymic, chemical, light microscopic, and electron microscopic examination showed less than 2% contamination by stromal contents, no contamination by microbial, microsomal, or mitochondrial membranes, and possible low levels of lamellar membrane contamination. Yields of 0.5 mg of envelope membrane protein were obtained from 56-g leaf sections. The Mg2+-dependent nonlatent ATPase activity, a marker enzyme for chloroplast envelope membranes, was 40 μmoles Pi released hr−1 mg protein−1, a value similar to that obtained with pure mesophyll chloroplast envelope membranes from other plants.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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