植物生理

Changes in the concentration of cytoplasmic calcium, cyt are central regulators in many cellular signal transduction pathways including many lipid-mediated regulatory networks. Given this central role that has during plant growth, monitoring spatial and temporal dynamics can ...

The discovery of the green fluorescent protein has revolutionized cell biology as it allowed researchers to visualize dynamic processes in living cells. The fusion of fluorescent protein variants with lipid binding domains that bind to specific phospholipids have been very instru ...

Lipid affinity beads can be used to identify novel proteins with lipid binding capacity or to determine binding prerequisites of known lipid-binding proteins. Here we describe several applications for which this tool can be used and which considerations have to be taken into account. In add ...

Protein–lipid interactions play an important role in cellular protein relocation, activation and signal transduction. The liposome-binding assay is a simple and inexpensive method to examine protein–lipid binding in vitro. The phospholipids used for liposome production are d ...

Protein–lipid interactions play an important role in lipid metabolism, membrane trafficking and cell �signaling by regulating protein localization, activation, and function. The Fat Blot assay is a relatively simple and inexpensive method to examine these interactions using n ...

Phospholipases of the A type constitute a large family of esterases that catalyze the hydrolysis of the fatty acid ester bonds in phospholipids and thus generate lysophospholipids and fatty acids. Both products or their metabolites are important signal molecules in the cellular adapt ...

Phosphatidic acid phosphatase (PAP; EC 3.1.3.4) catalyzes the dephosphorylation of phosphatidic acid (PA) to produce diacylglycerol (DAG) and inorganic phosphate. In seed plants, PA plays pivotal roles both as a precursor to membrane lipids and as a signaling molecule. As more informati ...

Phospholipase D (PLD) hydrolyzes structural phospholipids like phosphatidylcholine (PC) and phosphatidylethanolamine (PE) into phosphatidic acid (PA) and free choline/ethanolamine. In plants, this activity can be stimulated by a wide variety of biotic and abiotic stresses ...

Over the past decade, tremendous progress has been made toward understanding the physiological functions of individual members of the diverse phospholipase D (PLD) family of enzymes in plants. For instance, the involvement of plant PLD members has been shown or suggested in a wide variety of t ...

Plant nonspecific phospholipase C (NPC) is a recently described enzyme which plays a role in membrane rearrangement during phosphate starvation. It is also involved in responses of plants to brassinolide, abscisic acid (ABA), elicitors, and salt. The NPC activity is decreased in cells tre ...

Measurement of phospholipase C (PLC) activity in vitro is a valuable biochemistry technique easily applicable in samples from different organisms. It quantifies the enzymatic activity of a key protein involved in critical developmental functions in organisms such as plants, anim ...

One critical aspect of phosphoinositide signaling is the turnover of signaling molecules in the pathway. These signaling molecules include the phosphatidylinositol phosphates (PtdInsPs) and inositol phosphates (InsPs). The enzymes that catalyze the breakdown of these mole ...

Inositol lipid kinases are perhaps the easiest and most straightforward enzymes in the phosphoinositide pathway to analyze. In addition to monitoring lipid kinase-specific activity, lipid kinase assays can be used to quantify the inositol lipids present in isolated membranes (Jo ...

Phosphatidylinositol (PtdIns) synthase is a lipid-synthesizing enzyme responsible for the synthesis of the phospholipid, PtdIns. Its enzymatic properties have been studied in in vitro assays using either membrane-enriched fractions or the purified protein in reconstitut ...

Plant phospholipids and glycolipids can be analyzed by direct infusion electrospray ionization triple-quadrupole mass spectrometry. A biological extract is introduced in solvent by continuous infusion into the mass spectrometer’s electrospray ionization source, where ...

Galactolipids represent the predominant membrane lipid class in plants. In general, galactolipids are restricted to plastids, but during phosphate deficiency, they also accumulate in extraplastidial membranes. Two groups of plants can be distinguished based on the presence of a s ...

Biological lipid extracts often contain small amounts of lysophospholipids (LPLs). Since different functions are emerging for LPLs in lipid metabolism and signalling, there is need for a reliable and cost-effective method for their identification. For this purpose, authentic LPL s ...

In plants, phosphatidic acid (PA) functions as a metabolic precursor in the biosynthesis of glycerolipids, but it also acts as a key signaling lipid in the response to environmental stress conditions (Testerink and Munnik, J Exp Bot 62:2349–2361, 2011). In vivo 32P-radiolabeling assays have s ...

Diacylglycerol (DAG) is an important intermediate of lipid metabolism and a component of phospholipase C signal transduction. Quantification of DAG in plant membranes represents a challenging task because of its low abundance. DAG can be measured by direct infusion mass spectromet ...

The phosphoinositide signaling pathway is important for plant responses to many different stresses. As part of the responses to a stimulus, InsP3 levels may increase rapidly and transiently. The receptor binding assay for InsP3 described here is easy to use and an ideal method to monitor and co ...