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采用双向凝胶电泳技术分析叶片蛋白质：以核酮糖二磷酸羧化酶/加氧酶降解为例分析蛋白酶的作用，以及在提取过程中避免蛋白水解的方法。

Analysis of Leaf Proteins by Two-Dimensional Gel Electrophoresis: Protease Action as Exemplified by Ribulose Bisphosphate Carboxylase/ Oxygenase Degradation and Procedure to Avoid Proteolysis during Extraction.

机构信息

Laboratoire de Génétique des Systèmes Végétaux, G.I.S. Moulon, 91190 Gif-sur-Yvette, France.

出版信息

Plant Physiol. 1985 May;78(1):178-82. doi: 10.1104/pp.78.1.178.

DOI:10.1104/pp.78.1.178
PMID:16664194
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1064698/
Abstract

A previous genetic analysis by two-dimensional electrophoresis of wheat leaf proteins led us to hypothesize that several polypeptides were degradation products of the large subunit of ribulose bisphosphate carboxylase/oxygenase (EC 4.1.1.39).VARIOUS EXTRACTION PROCEDURES INCLUDING THE USE OF PROTEINASE INHIBITORS ALLOWED US TO: (a) confirm that the suspected polypeptides are indeed degradation products; (b) find out under which conditions the proteinases act; (c) find a method which prevents degradation, by boiling the extract in the presence of sodium dodecyl sulfate and 2-mercaptoethanol.

摘要

先前通过二维电泳对小麦叶片蛋白的遗传分析使我们假设，有几个多肽是核酮糖二磷酸羧化酶/加氧酶（EC 4.1.1.39）大亚基的降解产物。各种提取方法，包括使用蛋白酶抑制剂，使我们能够：（a）确认怀疑的多肽确实是降解产物；（b）找出蛋白酶在何种条件下起作用；（c）找到一种方法，即在十二烷基硫酸钠和 2-巯基乙醇存在下煮沸提取物，以防止降解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9b/1064698/592fe6ce0613/plntphys00588-0190-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9b/1064698/d926ca0bc43d/plntphys00588-0187-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9b/1064698/592fe6ce0613/plntphys00588-0190-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9b/1064698/d926ca0bc43d/plntphys00588-0187-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d9b/1064698/592fe6ce0613/plntphys00588-0190-a.jpg

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本文引用的文献

1
Source of endoproteolytic activity associated with purified ribulose bisphosphate carboxylase.与纯化的核酮糖二磷酸羧化酶相关的内肽酶活性的来源。
Plant Physiol. 1984 May;75(1):74-7. doi: 10.1104/pp.75.1.74.
2
Two-Dimensional Electrophoretic Mapping of Proteins of Bundle Sheath and Mesophyll Cells of the C(4) Grass Digitaria sanguinalis (L.) Scop. (Crabgrass).C4 禾本科植物马唐（Digitaria sanguinalis (L.) Scop.）维管束鞘细胞和叶肉细胞蛋白质的二维电泳图谱
Plant Physiol. 1982 Nov;70(5):1359-66. doi: 10.1104/pp.70.5.1359.
3
Partial purification and characterization of endoproteinases from senescing barley leaves.
利用双端体系的 2D 电泳技术对小麦结构基因和调控因子进行染色体定位。
Theor Appl Genet. 1985 Nov;71(1):31-8. doi: 10.1007/BF00278250.
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Is the polymorphism of protein amounts related to phenotypic variability? A comparison of two-dimensional electrophoresis data with morphological traits in maize.蛋白质数量的多态性与表型变异性有关吗？玉米中二维电泳数据与形态特征的比较。
Theor Appl Genet. 1987 Jun;74(2):194-202. doi: 10.1007/BF00289968.
5
Regulatory effects of homoeologous chromosome arms on wheat proteins at two developmental stages.同源染色体臂对两个发育阶段小麦蛋白的调控作用。
Theor Appl Genet. 1986 Dec;73(2):246-51. doi: 10.1007/BF00289281.
6
Proteases of Melilotus alba mesophyll protoplasts : II. General properties and effectiveness in degradation of cytosolic and vacuolar enzymes.白车轴草叶肉原生质体蛋白酶：Ⅱ. 一般性质及对胞质和液泡酶的降解效果。
Planta. 1987 Apr;170(4):541-9. doi: 10.1007/BF00402989.
7
Protein mapping and genome expression variations in the basidiomycete Agrocybe aegerita.担子菌阿魏侧耳的蛋白质图谱和基因组表达变化。
Theor Appl Genet. 1989 Oct;78(4):505-12. doi: 10.1007/BF00290834.
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Floral-specific polypeptides of the Japanese morning glory.日本牵牛花花器官特异性多肽。
Planta. 1988 Aug;175(2):221-8. doi: 10.1007/BF00392431.
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Patterns of protein synthesis in dormant and growing vegetative buds of pea.豌豆休眠和生长营养芽中的蛋白质合成模式。
Planta. 1988 Dec;176(4):497-505. doi: 10.1007/BF00397656.
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Ectomycorrhizins - symbiosis-specific or artifactual polypeptides from ectomycorrhizas?外生菌根共生多肽——外生菌根中的共生特异性或人为多肽？
Planta. 1992 Mar;188(1):129-36. doi: 10.1007/BF01160722.
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Plant Physiol. 1981 Oct;68(4):930-6. doi: 10.1104/pp.68.4.930.
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Plant Physiol. 1979 Nov;64(5):884-7. doi: 10.1104/pp.64.5.884.
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Breakdown of Ribulose Bisphosphate Carboxylase and Change in Proteolytic Activity during Dark-induced Senescence of Wheat Seedlings.小麦幼苗暗诱导衰老过程中核酮糖二磷酸羧化酶的降解及蛋白水解活性的变化
Plant Physiol. 1978 Oct;62(4):604-8. doi: 10.1104/pp.62.4.604.
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Proteases of Senescing Oat Leaves: II. Reaction to Substrates and Inhibitors.衰老燕麦叶片中的蛋白酶：II. 对底物和抑制剂的反应。
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A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels.一种用于检测聚丙烯酰胺凝胶中蛋白质的简化超灵敏银染法。
Anal Biochem. 1980 Jul 1;105(2):361-3. doi: 10.1016/0003-2697(80)90470-4.
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Studies on human plasma alpha 2-macroglobulin-enzyme interactions. Evidence for proteolytic modification of the subunit chain structure.人血浆α2-巨球蛋白与酶相互作用的研究。亚基链结构蛋白水解修饰的证据。
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High resolution two-dimensional electrophoresis of proteins.蛋白质的高分辨率二维电泳
J Biol Chem. 1975 May 25;250(10):4007-21.