EVIDENCE FOR SEPARATE SYSTEMS FOR THE TRANSPORT OF NEUTRAL AND BASIC AMINO ACIDS ACROSS THE BLOOD-BRAIN BARRIER

Abstract— The effects of high circulating concentrations of several amino acids on the free amino acids of rat brain were measured, to see whether or not the results followed any consistent pattern. High circulating concentrations of large, neutral amino acids (phenylalanine, valine or isoleucine) caused significantly decreased values only of other large, neutral amino acids in the brains. High circulating concentrations of the basic amino acids lysine or arginine caused significantly decreased values only of each other. The data suggest that there are separate systems for the transport of neutral and basic amino acids across the blood-brain barrier. The effects of valine and lysine on the uptake by brain and the con-vulsant action of allylglycine (a neutral amino acid) were consistent with the concept of separate systems for the transport of amino acids across the blood-brain barrier. Valine inhibited the uptake by brain and the convulsant action of allylglycine in mice, but lysine did not. The data suggest that allylglycine and valine are transported into the brain by a common mechanism that does not transport lysine.     

Transport Processes in Vacuoles of Higher Plants

The vacuole is the largest compartment of a mature plant cell and serves as an internal reservoir of metabolites and nutrients. In the last years transport of solutes across the tonoplast has been intensively investigated. It was shown that two different proton pumps reside in the tonoplast. These pumps generate an electrochemical gradient which can be used as an energy-source to accumulate solutes. Cation uptake is driven by an H⁺ antiport mechanism. Anions are accumulated in response to the inside positive membrane potential. In addition, the existence of ion channels was shown using the patch clamp technique. The aim of this review is to compare and to discuss the present state of our knowledge of solute transport across the tonoplast.     

高等植物蔗糖转运的分子调控

在高等植物中,蔗糖的合成、运输与分配是一个复杂的过程。蔗糖由源到库的运输不仅与植物的生长发育相关,还受到植物体内的激素水平以及外界环境条件变化等因素的影响。蔗糖转运蛋白介导了蔗糖在植物韧皮部的装载、运输和卸载,在某些库中的蔗糖转运和库组织分配的分子调控中起有重要的生理作用。此外,简要介绍了笔者实验室在橡胶树蔗糖转运蛋白基因研究方面的最新进展。     

高等植物钾转运蛋白

钾在植物生长发育过程中具有许多重要的作用。以模式植物拟南芥中克隆和鉴定的钾通道和转运体为基础,全面介绍了高等植物中钾转运体系家族,包括Shaker通道、KCO通道、KUP/HAK/KT转运体、HKT转运体和其它转运体。同时,分析了在高等植物中存在多种钾吸收和转运机制的可能原因。     

The Transport and Metabolism of Urea in Chara australis: III. TWO SPECIFIC TRANSPORT SYSTEMS

Reciprocal competitive inhibition studies were used to showthat N-methyl-urea (NMU), acetamide and urea all compete forbinding to a common transport system, designated system I andthat this system is one of two specific mechanisms transportingurea in Chara. System I binds urea with a Km of about 0–3mmol m-3 and is strongly influenced by metabolic controls. SystemI is active and electrogenic and may be energized by the couplingof urea uptake to an influx of protons. This is the first reportof an electrogenic urea transport system in an alga. The secondspecific mechanism for urea transport, designated system II,binds urea with a relatively low affinity (K_m c. 7–0 mmolm-3) and does not transport NMU to a significant extent. SystemII is less subject to metabolic control than system I and, thoughit may be active, is not electrogenic. Key words: Urea, methylurea, proton cotransport, metabolic control     

Auxin Transport Inhibitors: IV. EVIDENCE OF A COMMON MODE OF ACTION FOR A PROPOSED CLASS OF AUXIN TRANSPORT INHIBITORS: THE PHYTOTROPINS

The more active members of a proposed class of auxin transport inhibitors have been shown to have the ability to inhibit the active movement of auxin at concentrations where they have little effect on auxin action and no significant auxin activity. They have also been shown to give rise to characteristic biphasic dose-response curves on cress root growth. Based on these physiological similarities and other common physiological properties, it is concluded that they may achieve their effects by a common mode of action which differs from that of other known auxin transport inhibitors. It is suggested that the name "phytotropins" be given to the class of auxin transport inhibitors now defined by a similar mode of action and common chemical properties.     

高等植物尿素代谢及转运的分子机理

尿素广泛存在于自然界中,是易于被许多生物（如植物）利用的生长氮源。该文通过概述尿素在不同生命系统中存在的基础生理意义及各类型尿素转运蛋白,讨论了植物细胞中尿素合成与分解的各种途径及尿素在植物氮营养、代谢和运输中的生理作用。迄今为止,在植物中已发现了2类转运尿素的膜蛋白,即MIPs和DUR3,它们分别在低亲和力、高亲和力尿素运输中发挥潜在作用。异源表达结果表明MIPs介导了尿素的被动迁移：而AtDUR3则参与拟南芥根系对尿素的吸收。对MIPs和DUR3转运尿素的酶学特征、亚细胞作用位点和表达调控状况等的研究表明：它们的分子生物学功能与植物的氮营养及氮素再分配和利用相关。     

Inhibition of Amino Acid Transport in Rabbit Intestine by p-Chloromercuriphenyl Sulfonic Acid

Influx of phenylalanine across the brush border of rabbit intestine is markedly reduced by treatment with 5 mM p-chloromercuriphenyl sulfonate (PCMBS). The effect is rapidly and completely reversed by dithiothreitol. Phenylalanine influx into PCMBS-treated tissue can be competitively inhibited by other neutral amino acids and follows saturation kinetics. PCMBS causes an increase in the apparent Michaelis constant from the value observed in control tissue but does not alter the maximal influx significantly. Treatment of the tissue with PCMBS leads to a significant reduction in the Na-sensitivity of the transport, and a number of results indicate that the major effect of the reagent is to cause a marked reduction in the affinity of the transport system for Na. The transport system can be partially protected against reaction with PCMBS by phenylalanine and tryptophan but not by methionine or norleucine. The results suggest that PCMBS reacts with a sulfhydryl group in the region of the transport site and may alter conformational changes associated with the binding of substrates.     

高等植物尿素代谢及转运的分子机理

尿素广泛存在于自然界中, 是易于被许多生物(如植物)利用的生长氮源。该文通过概述尿素在不同生命系统中存在的基础生理意义及各类型尿素转运蛋白, 讨论了植物细胞中尿素合成与分解的各种途径及尿素在植物氮营养、代谢和运输中的生理作用。迄今为止, 在植物中已发现了2类转运尿素的膜蛋白, 即MIPs和DUR3, 它们分别在低亲和力、高亲和力尿素运输中发挥潜在作用。异源表达结果表明, MIPs介导了尿素的被动迁移; 而AtDUR3则参与拟南芥根系对尿素的吸收。对MIPs和DUR3转运尿素的酶学特征、亚细胞作用位点和表达调控状况等的研究表明: 它们的分子生物学功能与植物的氮营养及氮素再分配和利用相关。     

Transport of Salicylic Acid in Tobacco Necrosis Virus-Infected Cucumber Plants

The transport of salicylic acid (SA) was studied in cucumber (Cucumis sativus L.) using 14C-labeled benzoic acid that was injected in the cotyledons at the time of inoculation. Primary inoculation with tobacco necrosis virus (TNV) on the cotyledons led to an induction of systemic resistance of the first primary leaf above the cotyledon against Colletotrichum lagenarium as early as 3 d after inoculation. [14C]SA was detected in the phloem or in the first leaf 2 d after TNV inoculation, whereas [14C]benzoic acid was not detected in the phloem during the first 3 d after TNV inoculation of the cotyledons, indicating phloem transport of [14C]SA from cotyledon. In leaf 1, the specific activity of [14C]SA decreased between 1.7 and 8.6 times compared with the cotyledons, indicating that, in addition to transport, leaf 1 also produced more SA. The amount of SA transported after TNV infection of the cotyledon was 9 to 160 times higher than in uninfected control plants. Thus, SA can be transported to leaf 1 before the development of systemic acquired resistance, and SA accumulation in leaf 1 results both from transport from the cotyledon and from synthesis in leaf 1.     

Intercellular Transport in Plants: I. THE RATE OF TRANSPORT OF CHLORIDE AND THE ELECTRIC RESISTANCE

A quantitative study has been made of the intercellular movementof chloride in Chara corallina, using pairs of joined internodalcells. One cell of the pair (cell 1) was exposed to a solutioncontaining ³⁶Cl; the distribution of this tracer between thecells was determined at the end of the uptake period. Of thechloride taken up, 0.29 was transported out of cell 1 for alluptake times from 1.8 to 22 ks and 0.57 was transported to thevacuole of cell 1, in experimental series I. In series II, thefraction transported out of cell 1 was 0.43 at 22 and 43 ks,but 0.17 at 600 s. These results represent a rate of transport of 4 to 60 pmols^–1, across an intercellular wall of area 1.5 x 10^–6m²; the wall has 0.03 to 0.04 of its area occupied by plasmodesmata.The estimate of transport rate is based on an attempt to determinethe specific activity of the cytoplasm of cell 1. The electricresistance of the node was found to be 47 m m². The observed transport rate can be explained by diffusion inthe plasmodesmata, without the need to postulate active processes.Diffusion in the plasmodesmata is slower than in free solutionby an ‘impediment factor’ of 7 to 700, dependingon the assumed chloride concentration of the ground-plasm. Ifthe plasmodesmata offer the major conducting path for electriccurrent, the electric impediment factor is 390. Chloride entersthe plasmodesmata from the same small kinetic compartment whichsupplies the flux to the vacuole, or from a smallintermediatecompartment.     

{gamma}-Aminobutyric Acid Metabolism in Plants: Part 2. Metabolism in Higher Plants

The metabolism of -aminobutyric acid (AB) has been studied inhigher plants, particularly in peas and peanuts. Transaminationappeared to form the first step in AB degradation although transaminaseactivities were very low. The relatively active AB transaminaseassociated with whole pea plants possessing nodulated rootsappears to reside almost entirely within the nodules. AB transaminationwas demonstrated conclusively in extracts of mitochondria fromcotyledons of peanut seedlings; pyruvic acid acted as a betteramino-group acceptor than -ketoglutaric acid (KG). AB transaminaseactivity present in the microsomal and soluble cytoplasmic fractionsof the cells was very low AB was not metabolized perceptibly by intact mitochondria frompeanut, but when various organic acids were supplied simultaneously,an extra uptake of oxygen occurred and was associated with ABdisappearance. Aspartate, alanine, and ammonia were formed usingthe nitrogen atom of AB. The metabolic pathway followed by the carbon skeleton of ABwas traced by supplying C¹⁴-labelled material to leaf discsof peas and to mitochondria from peanut cotyledons. Radioactivitywas incorporated into organic acids, amino-acids, and respiratorycarbon dioxide in a manner suggesting that AB was convertedinto succinate which was then metabolized by the enzymes ofthe Krebs cycle present in the plant mitochondria. Glutamic decarboxylase was shown to be present largely in thenon-particulate (soluble) cytoplasm of cells. The enzymes responsiblefor AB synthesis and degradation, glutamic decarboxylase, andAB transaminase, respectively, therefore largely reside in differentsub-cellular fractions.     

Intercellular Transport in Plants: II. CYCLOSIS AND THE RATE OF INTERCELLULAR TRANSPORT OF CHLORIDE IN CHARA

The effect of streaming speed on intercellular transport ofchloride has been studied using pairs of internodal cells ofChara. The rate of transport was measured by that fraction ofthe chloride that entered one internode which was transportedout of it into the cells of the node and the next internode.The speed of cytoplasmic streaming was altered by treating thefirst cell with cytochalasin B. The relative rate of intercellular transport depended markedlyon the streaming speed at all speeds up to those found in untreatedcells. The chloride influx into the treated cell did not dependon the streaming speed. It is concluded that the rate of intercellular transport oflow molecular weight solutes in Chara will be normally limitedby the rate at which cytoplasmic streaming brings solute tothe plasmodesmata, rather than by the diffusion permeabilityof the plasmodesmata. This conclusion may well apply to othercharophyte plants, and could in principle apply to higher plants.     

Angiopoietin-like Protein 4 Inhibition of Lipoprotein Lipase: EVIDENCE FOR REVERSIBLE COMPLEX FORMATION*

Elevated triglycerides are associated with an increased risk of cardiovascular disease, and lipoprotein lipase (LPL) is the rate-limiting enzyme for the hydrolysis of triglycerides from circulating lipoproteins. The N-terminal domain of angiopoietin-like protein 4 (ANGPTL4) inhibits LPL activity. ANGPTL4 was previously described as an unfolding molecular chaperone of LPL that catalytically converts active LPL dimers into inactive monomers. Our studies show that ANGPTL4 is more accurately described as a reversible, noncompetitive inhibitor of LPL. We find that inhibited LPL is in a complex with ANGPTL4, and upon dissociation, LPL regains lipase activity. Furthermore, we have generated a variant of ANGPTL4 that is dependent on divalent cations for its ability to inhibit LPL. We show that LPL inactivation by this regulatable variant of ANGPTL4 is fully reversible after treatment with a chelator.     

Metabolism of 2,4-Dichlorophenoxyacetic Acid (2,4-D) in Soybean Root Callus : EVIDENCE FOR THE CONVERSION OF 2,4-D AMINO ACID CONJUGATES TO FREE 2,4-D

An auxin-requiring soybean root callus metabolized [1-¹⁴C]-2,4-dichlorophenoxyacetic acid (2,4-D) to diethyl ether-soluble amino acid conjugates and water-soluble metabolites. The uptake in tissue varied with incubation time, concentration, and amount of tissue. Uptake was essentially complete (80%) after a 24-hour incubation and the percentage of free 2,4-D in the tissue fell to its lowest point at this time. At later times, the percentage of free 2,4-D increased and the percentage of amino acid conjugates decreased, whereas the percentage of water-soluble metabolites increased only slightly. Similar trends were seen if the tissue was incubated for 24 hours in radioactive 2,4-D, followed by incubation in media without 2,4-D for 24 hours. Inclusion of nonlabeled 2,4-D during the 24-hour chase period did not reduce amino acid conjugate disappearance but did reduce the percentage of free [1-¹⁴C]2,4-D. Thus, an external supply of 2,4-D does not directly prevent amino acid conjugate metabolism in this tissue. It is concluded that 2,4-D amino acid conjugates were actively metabolized by this tissue to free 2,4-D and water-soluble metabolites.     

Effect of Thiolactomycin on Fatty Acid Synthesis in Higher Plants

Thiolactomycin (TLM), an antibiotic from Nocardia sp. No. 2-200,inhibited fatty acid synthesis in Avena leaves, with the concentrationcausing 50% inhibition being 0.38µg/ml. This antibioticis more inhibitory to the elongation of palmitic to oleic acidthan to the de novo synthesis of palmitic acid in both spinachchloroplasts and Avena leaves, in contrast to the effect ofcerulenin which inhibits de novo synthesis but not fatty acidelongation. On the other hand, TLM is less inhibitory to furtherelongation of stearic acid to very long chain fatty acids inpea seeds. The inhibition rate decreased in the order of synthesisof arachidic, behenic and lignoceric acid. (Received December 26, 1986; Accepted April 24, 1987)     

高等植物脱落酸的生物合成及其调控

介绍了近年来高等植物体内ABA的合成部位,ABA生物合成缺陷型突变体,ABA生物合成途径及其调控的最新研究进展。     

Inhibition of Axoplasmic Transport in the Optic System by Kainic Acid

Axoplasmic transport along the optic axons was studied after intraocular injections of kainic acid (KA). Transport of labeled material did not initiate from the eye when KA was injected simultaneously with the protein precursor [3H]proline. When KA was injected after axoplasmic transport of labeled proteins had begun, no additional radioactive material moved out of the retinal ganglion cells. However, the labeled material already present in the optic nerve at the time of KA injection continued to move, and accumulated at the nerve endings. Although KA reduces the incorporation of precursor, this effect of KA on axoplasmic transport appears to be more than a consequence of inhibition on precursor uptake or protein synthesis. Recovery from this KA action began 6 h after exposure to KA and was about 50% recovered by 36 h. The extent of the recovery remained at this level for as long as a week, which suggested a partial recovery of the ganglion cells. A second exposure to KA after the inner plexiform layer had virtually disappeared was as effective as the first exposure in preventing the appearance of transported protein in the optic nerve, suggesting a direct action of KA on the ganglion cells. We interpreted the results to indicate that KA interferes with the initiation phase of axoplasmic transport in ganglion cells and this effect is partially reversible.