A small scale procedure for the isolation of transport competent vesicles from plant tissues

John L. Giannini; Jose Ruiz-Cristin; Donald P. Briskin

doi:10.1016/0003-2697(88)90056-5

A small scale procedure for the isolation of transport competent vesicles from plant tissues

John L. Giannini, Jose Ruiz-Cristin, Donald P. Briskin

Crop Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

A microscale method for the isolation of selectively sealed microsomal membrane fractions from plant tissue is presented. The method is based on differential centrifugation in a table top microcentrifuge to accommodate small sample size (10-25 g tissue) and the addition of KI or KCl in the homogenization medium for isolating selectively sealed plasma membrane or tonoplast vesicles. This microscale procedure was found to be useful in isolating membranes from red beet (Beta vulgaris) storage tissue, sugar beet (Beta vulgaris) storage tissue, corn (Zea mays) roots, and soybean (Glycine max) roots. This paper also describes the ability to further purify an enriched red beet plasma membrane fraction on a discontinuous sucrose density gradient, in a microcentrifuge, that is highly competent in ATP-dependent H⁺-transport. The speed and wide applicability of this procedure make it ideal when a large number of samples need to be processed.

Original language	English (US)
Pages (from-to)	561-567
Number of pages	7
Journal	Analytical Biochemistry
Volume	174
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(88)90056-5
State	Published - Nov 1 1988

Keywords

H-transporting ATPases
membrane transport
plasma membrane
tonoplast
vesicle

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Online availability

10.1016/0003-2697(88)90056-5

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title = "A small scale procedure for the isolation of transport competent vesicles from plant tissues",

abstract = "A microscale method for the isolation of selectively sealed microsomal membrane fractions from plant tissue is presented. The method is based on differential centrifugation in a table top microcentrifuge to accommodate small sample size (10-25 g tissue) and the addition of KI or KCl in the homogenization medium for isolating selectively sealed plasma membrane or tonoplast vesicles. This microscale procedure was found to be useful in isolating membranes from red beet (Beta vulgaris) storage tissue, sugar beet (Beta vulgaris) storage tissue, corn (Zea mays) roots, and soybean (Glycine max) roots. This paper also describes the ability to further purify an enriched red beet plasma membrane fraction on a discontinuous sucrose density gradient, in a microcentrifuge, that is highly competent in ATP-dependent H+-transport. The speed and wide applicability of this procedure make it ideal when a large number of samples need to be processed.",

keywords = "H-transporting ATPases, membrane transport, plasma membrane, tonoplast, vesicle",

author = "Giannini, {John L.} and Jose Ruiz-Cristin and Briskin, {Donald P.}",

note = "Funding Information: {\textquoteright} This research was supported by USDA Competitive Grant No. 86-I-CRCR-1977 and a McKnight Foundation Individual Research Award in Plant Biology awarded to D.P.B. {\textquoteright} Permanent address: Department of Biology, St. Olaf College, Northfield, MN 55057.",

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doi = "10.1016/0003-2697(88)90056-5",

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AU - Giannini, John L.

AU - Ruiz-Cristin, Jose

AU - Briskin, Donald P.

N1 - Funding Information: ’ This research was supported by USDA Competitive Grant No. 86-I-CRCR-1977 and a McKnight Foundation Individual Research Award in Plant Biology awarded to D.P.B. ’ Permanent address: Department of Biology, St. Olaf College, Northfield, MN 55057.

PY - 1988/11/1

Y1 - 1988/11/1

N2 - A microscale method for the isolation of selectively sealed microsomal membrane fractions from plant tissue is presented. The method is based on differential centrifugation in a table top microcentrifuge to accommodate small sample size (10-25 g tissue) and the addition of KI or KCl in the homogenization medium for isolating selectively sealed plasma membrane or tonoplast vesicles. This microscale procedure was found to be useful in isolating membranes from red beet (Beta vulgaris) storage tissue, sugar beet (Beta vulgaris) storage tissue, corn (Zea mays) roots, and soybean (Glycine max) roots. This paper also describes the ability to further purify an enriched red beet plasma membrane fraction on a discontinuous sucrose density gradient, in a microcentrifuge, that is highly competent in ATP-dependent H+-transport. The speed and wide applicability of this procedure make it ideal when a large number of samples need to be processed.

AB - A microscale method for the isolation of selectively sealed microsomal membrane fractions from plant tissue is presented. The method is based on differential centrifugation in a table top microcentrifuge to accommodate small sample size (10-25 g tissue) and the addition of KI or KCl in the homogenization medium for isolating selectively sealed plasma membrane or tonoplast vesicles. This microscale procedure was found to be useful in isolating membranes from red beet (Beta vulgaris) storage tissue, sugar beet (Beta vulgaris) storage tissue, corn (Zea mays) roots, and soybean (Glycine max) roots. This paper also describes the ability to further purify an enriched red beet plasma membrane fraction on a discontinuous sucrose density gradient, in a microcentrifuge, that is highly competent in ATP-dependent H+-transport. The speed and wide applicability of this procedure make it ideal when a large number of samples need to be processed.

KW - H-transporting ATPases

KW - membrane transport

KW - plasma membrane

KW - tonoplast

KW - vesicle

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A small scale procedure for the isolation of transport competent vesicles from plant tissues

Abstract

Keywords

ASJC Scopus subject areas

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