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来自玉米（Zea mays L.）胚芽鞘的两种膜相关β-葡萄糖苷酶的特性分析。

Characterization of two membrane-associated beta-glucosidases from maize (Zea mays L.) coleoptiles.

作者信息

Feldwisch J, Vente A, Zettl R, Bako L, Campos N, Palme K

机构信息

Max-Planck Institut für Züchtungsforschung, Köln, Federal Republic of Germany.

出版信息

Biochem J. 1994 Aug 15;302 ( Pt 1)(Pt 1):15-21. doi: 10.1042/bj3020015.

DOI:10.1042/bj3020015
PMID:8068000
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1137185/
Abstract

We isolated membrane vesicles from maize (Zea mays L.) coleoptiles and identified in these vesicles a 58 kDa (pm58) and a 60 kDa (pm60) protein by photoaffinity labelling with 5-azido-[7-3H]indole-3-acetic acid ([3H]N3IAA). Photoaffinity labelling was effectively competed for by auxins as well as by flavonoids. The labelled proteins were solubilized by Triton X-114 from the vesicles and partially purified. Microsequence analysis revealed that pm60 is a beta-glucosidase. This was confirmed by biochemical and immunological analysis. We show that pm60 has a beta-D-glucoside glucohydrolase (EC 3.2.1.21) activity. It uses p-nitro-phenyl beta-D-glucopyranoside (PNPG) as a substrate, with a pH optimum of 5.0. The Km for PNPG is 0.652 mM and the Vmax. 6.24 mumol.min-1.mg-1. The beta-glucosidase activity of pm60 was competitively inhibited by IAA and 1-naphthylacetic acid as well as by gluconolactam and glucose. N-terminal amino-acid-sequence analysis of pm58 revealed similarity to pm60, suggesting that both proteins are encoded by different members of a gene family.

摘要

我们从玉米（Zea mays L.）胚芽鞘中分离出膜泡，并通过用5-叠氮基-[7-³H]吲哚-3-乙酸（[³H]N3IAA）进行光亲和标记，在这些膜泡中鉴定出一种58 kDa（pm58）和一种60 kDa（pm60）的蛋白质。生长素以及类黄酮能有效竞争光亲和标记。标记的蛋白质用Triton X-114从膜泡中溶解并部分纯化。微序列分析表明pm60是一种β-葡萄糖苷酶。这通过生化和免疫分析得到证实。我们发现pm60具有β-D-葡萄糖苷葡萄糖水解酶（EC 3.2.1.21）活性。它以对硝基苯基β-D-吡喃葡萄糖苷（PNPG）为底物，最适pH为5.0。PNPG的Km为0.652 mM，Vmax为6.24 μmol·min⁻¹·mg⁻¹。pm60的β-葡萄糖苷酶活性受到吲哚乙酸、1-萘乙酸以及葡糖醛酸内酯和葡萄糖的竞争性抑制。pm58的N端氨基酸序列分析显示与pm60相似，表明这两种蛋白质由一个基因家族的不同成员编码。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/0200548780c4/biochemj00081-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/7e04c44e3f7e/biochemj00081-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/02b710ecb1fd/biochemj00081-0026-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/481cd1193bdc/biochemj00081-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/0200548780c4/biochemj00081-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/7e04c44e3f7e/biochemj00081-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/02b710ecb1fd/biochemj00081-0026-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/481cd1193bdc/biochemj00081-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e704/1137185/0200548780c4/biochemj00081-0028-a.jpg

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