The effect of nitrate, nitrite, and ammonia on photosynthesis by Acetabularia chloroplast preparations compared with spinach chloroplasts and whole cells of Acetabularia and Dunaliella

1972 ◽  
Vol 50 (12) ◽  
pp. 2535-2543 ◽  
Author(s):  
B. R. Grant ◽  
F. Winkenbach ◽  
D. T. Canvin ◽  
R. G. S. Bidwell

The effects of nitrate, nitrite, and ammonia were compared using chloroplasts prepared from Acetabularia mediterranea and spinach and intact cells of Acetabularia and Dunaliella tertiolecta. Nitrate was without effect in all systems. Spinach chloroplasts were strongly inhibited by nitrite at 5 mM, Acetabularia cells were somewhat inhibited, but Dunaliella cells and chloroplast preparations from Acetabularia were unaffected. Ammonia at 5 mM inhibited spinach and Acetabularia chloroplasts, but its effect on the other systems was slight. It decreased oxygen production in chloroplasts and Dunaliella cells, but did not affect carbon dioxide fixation.These results are interpreted on the basis of varying rates of uptake, metabolism, and detoxification of the ions in different preparations. Whole cells and Acetabularia chloroplast preparations make amino acids, but spinach chloroplasts do not. This accounts for the greater sensitivity of the latter, and for the decreased oxygen production in the presence of ammonia in whole cells, which is presumably caused by increased respiration accompanying amino acid formation. The insensitivity of the Acetabularia chloroplast preparation is attributed to the properties of the membrane, apparently derived from tonoplast fragments, which surrounded the small cytoplasmic droplets present in these preparations.

1972 ◽  
Vol 50 (12) ◽  
pp. 2545-2551 ◽  
Author(s):  
F. Winkenbach ◽  
B. R. Grant ◽  
R. G. S. Bidwell

Nitrogen was supplied in the form of nitrate, nitrite, or ammonium ions to whole cells of Dunaliella tertiolecta, to whole cells and chloroplast preparations of Acetabularia mediterranea, and to spinach chloroplasts, while they were photosynthesizing in 14CO2. The 14C labeling patterns in these experiments provide information on several aspects of the photosynthesis, nitrogen metabolism, and physical properties of these systems. The rates and products of photosynthesis are affected in different ways by different ions, depending on the penetration of each ion, its toxicity, and on the ability of the system under test to synthesize amino acids. Thus the ions that penetrate spinach chloroplasts, while they may inhibit photosynthesis, do not affect the distribution of 14C among photosynthetic products because no amino acids are formed. The large differences in behavior between Acetabularia whole cells and chloroplast preparations from these cells suggest that the membrane structure surrounding cytoplasmic droplets in the latter may be tonoplast rather than plasmalemma in origin.


1976 ◽  
Vol 154 (1) ◽  
pp. 43-48 ◽  
Author(s):  
J D Young ◽  
J C Ellory ◽  
E M Tucker

1. Uptake rates for 23 amino acids were measured for both normal (high-GSH) and GSH-deficient (low-GSH) erythrocytes from Finnish Landrace sheep. 2. Compared with high-GSH cells, low-GSH cells had a markedly diminished permeability to D-alanine, L-alanine, α-amino-n-butyrate, valine, cysteine, serine, threonine, asparagine, lysine and ornithine. Smaller differences were observed for glycine and proline, whereas uptake of the other amino acids was not significantly different in the two cell types.


1963 ◽  
Vol 53 (4) ◽  
pp. 681-713 ◽  
Author(s):  
R. G. Fennah

The feeding of the cacao thrips, Selenothrips rubrocinctus (Giard), on cashew, Anacardium occidentale, one of its host plants in Trinidad, West Indies, is considered in relation to the annual period of maximum population increase on this host and to the choice of feeding sites on individual leaves. On trees observed for three years, populations regularly increased during the dry season, from a low level in December and January to a peak in April or May, and then rapidly declined during the wet season. Even when thrips were most abundant, some trees were free from attack, and this could not be attributed to protective morphological features, to specific repellent substances in the leaf, or to chance. S. rubrocinctus was found to feed on leaves that were subjected to water-stress and to breed only on debilitated trees: the evidence suggested that the adequacy of its supply of nutrients depends on the induction of suitable metabolic conditions within the leaf by water-stress.Both nymphs and adults normally feed on the lower, stomata-bearing surface of the leaf, but in a very humid atmosphere only a weak preference is shown for this surface and if, under natural conditions, it is exposed to insolation by inversion of the leaf, the insects migrate to the other surface. Since the thrips were shown to be indifferent to bodily posture, the observation suggests that their behaviour is governed primarily by avoidance of exposure to undue heat or dryness and only secondarily by the attractiveness of the stomata-bearing surface.Leaves of cashew tend not to become infested while still immature, and become most heavily infested, if at all, soon after they have hardened. Breeding does not occur on senescent leaves. The positions of feeding thrips are almost random on leaves under abnormal water-stress, but otherwise conform to certain patterns that mainly develop in fixed sequence. On reversal of an undetached leaf and consequent transfer of thrips from one surface to the other, there is no appreciable change in their distribution pattern or the apparent acceptability of the substrate. Changes of pattern were readily induced by injury to the plant during a period of water-stress and less easily, or not at all, when water-stress was low. Injury of areas of the leaf by heat was followed by their colonisation by thrips, and partial severance of branches by increased attack on their leaves.Leaves detached from uninfested trees invariably became acceptable for feeding within four hours. During this period, leaf water-content declined and the ratios of soluble-carbohydrate content and α-amino acids to fresh-leaf weight fell slightly and rose considerably, respectively. In the field, the latter ratio was invariably higher for infested than for uninfested leaf tissue, even on portions of the same leaf. If the nutrient value of leaf tissue is determined by the rate at which α-amino acids are extractable through a stylet puncture, the observed change in acceptability for feeding following plucking may be accounted for by the increase in α-amino-acid concentration. Feeding that is restricted on any one tree to the margins of local leaf injuries during prolonged high water-stress and totally absent when stress is low can be correlated with an α-amino-acid content in the living marginal tissue that is high or low, respectively. The ability of thrips to establish themselves and breed on leaves of a particular tree in the dry season and their failure to do so on leaves of the same tree in the wet season conforms with the greater or less amino-acid concentration occurring in the leaf at these respective times.


1981 ◽  
Vol 91 (3) ◽  
pp. 860-865 ◽  
Author(s):  
J J Rauh ◽  
D L Nelson

Extruded trichocysts are composed of a family of proteins with molecular weights between 15,000 and 20,000. We have used heat treatment and affinity chromatography on fluphenazine-Sepharose to purify calmodulinlike proteins from whole cells and from extruded trichocysts. The purified protein from trichocysts is indistinguishable from that of whole cells; it is heat-stable, activates brain phosphodiesterase in a Ca++-dependent fashion, changes mobility on SDS polyacrylamide gels in the presence of Ca++, contains 1 mol of trimethyllysine/17 kdaltons, and has the amino acid composition characteristic of calmodulins. Calmodulin is a major component of purified, extruded trichocysts, of which it represents between 1 and 10% by mass. The other proteins of the trichocyst also resemble calmodulin in several properties. Possible roles for calmodulin in the Ca++-activated extrusion of trichocysts is discussed.


1961 ◽  
Vol 39 (11) ◽  
pp. 1717-1735 ◽  
Author(s):  
P. G. Scholefield

The cumulative entry of amino acids into Ehrlich ascites carcinoma cells is due to the presence of active transport systems, each with its own specific range of substrates. Several amino acids and amino acid analogues may have an affinity for the same transport system and thus may inhibit transport of other amino acids by acting as competitive inhibitors or competitive substrates. Loss of methionine from ascites cells takes place by a diffusion process which obeys Fick's law. Leucine accumulation by ascites cells is small and is increased on addition of certain other amino acids. The increase is not due to inhibition of leucine oxidation as increase in the rate of production of radioactive carbon dioxide from labeled leucine also occurs. Kinetic aspects of these results are discussed.


Author(s):  
George Joseph ◽  
Asha Varughese ◽  
Ann Daniel

Abstract Background A method has been developed and validated for selective, accurate and precise determination of total proteinogenic amino acids and taurine from Infant Formula and Adult/Pediatric Nutritional Formulas (powders, ready-to-feed liquids, and liquid concentrates). The method was reviewed by the AOAC INTERNATIONAL SPIFAN Expert Review Panel (ERP) during the 133rd AOAC Annual Meeting & Expo on September 7, 2019 in Denver, CO, USA and was recommended to First Action Official MethodsSM status. Objective The method involves protein hydrolysis to amino acids, a simple pre-column derivatisation of amino acids and separation of derivatised amino acids by UHPLC. The quantification of amino acids is performed by multi-point calibration using norvaline as the internal standard. The analytical method is capable of quantitative determination for 22 proteinogenic amino acids, but cannot be used to quantitate tryptophan, which is destroyed during the acid hydrolysis step. Asparagine is determined as aspartic acid and glutamine as glutamic acid. The cystine and cysteine are converted to S-2-carboxyethylthiocysteine (CYSx) and the derivative is separated from the other amino acids. Citrulline which is present in some matrices and it is separated from other amino acids is not included in the method performance evaluation in the single laboratory validation (SLV). Method The proposed method met all the performance requirement limits set in standard method performance requirements (SMPR) 2014.013 for total proteinogenic amino acids and taurine. The correlation coefficient of multi-point calibration was not less than 0.999 for any amino acids at any point in the SLV study confirming the validity of linear dynanic range (LDR) and linearity of the method. The individual amino acids in the chromatogram were identified by absolute retention time and relative retention time (RRT) with respect to the internal standard norvaline. There were no significant (S/N Ratio <10) interferences from the reagents or by-products of derivatisation and targeted matrices. The method demonstrated high selectivity. Result Accuracy of the method was validated using standard reference materials (NIST SRM 1869 and 1849a) and spike recovery studies. The amino acid results in the SRMs were within the ranges of Reference Mass Fraction Values. The accuracy of the method was corroboratively validated by spike recovery studies. The average spike recovery range between 93 to 107% ensure the accuracy of the method for amino acids and compliance to the AOAC SMPR 2014.013. Conclusions Precision data of the method demonstrate that it meets the stakeholder requirements as per the SMPR. The mean RSDr for all the amino acids for 17 matrices selected for the SLV were not more than 4%. The method is very sensitive and the LOQ can go down to approximately ten times lower than the SMPR requirements. The sensitivity of method is a direct reflection of its signal to noise ratio which ensures guaranteed method performance at the lower levels of amino acids in these matrices. Highlights Taurine (aminoethane sulfonic acid) unlike the other amino acids is a beta-sulfonic amino acid that is not used in protein synthesis but is found as a free amino acid in tissues. The acidic functional group (-COOH) in common amino acid is replaced with a sulfonic acid (-SO3H) group in Taurine. The method offers baseline separation of citrulline which is an alpha amino acid generally present in Infant Formula and Adult/Pediatric Nutritional products. The separation of citrulline eliminates the risk of interference of this compound with other amino acids. The method can also separate and quantitate hydroxyproline, an important component of collagen that is often used to quantitate collagen. The method is simple and does not include any proprietary chemicals or instruments and can be performed on any basic reverse phase UHPLC system with UV detection.


1962 ◽  
Vol 40 (1) ◽  
pp. 459-469 ◽  
Author(s):  
P. H. Jellinck ◽  
Louise Irwin

Aerobic incubation of estrone-16-C14with peroxidase in the presence of serum albumin and other proteins resulted in the formation of water-soluble, ether-insoluble metabolites in high percentage yields. Similar products were formed when protein was replaced by cysteine or tryptophan but none of the other amino acids tested had any effect. The evidence points to an initial generation of hydrogen peroxide from these nitrogenous compounds by the enzyme acting as an aerobic oxidase, and the subsequent peroxidation of estrone to highly reactive products. These then combine with the protein or amino acid or else undergo alternative reactions. A strong chemical bond is formed with albumin and attempts to release the estrone metabolites from it were unsuccessful. Uterine homogenates from estrogen-treated rats showing high DPNH oxidase activity contained no "peroxidase" as measured by the formation of water-soluble products from estrone in the presence of protein.


1952 ◽  
Vol 30 (7) ◽  
pp. 522-528 ◽  
Author(s):  
W. B. McConnell

The carbon dioxide evolved when α-amino acids were heated for one hour at 85 °C. with ninhydrin was determined in a partially evacuated microdiffusion cell. Distillation of solvent from one chamber to the other was minimized by keeping the ionic concentrations of the reaction mixture and absorbing mixture approximately equal. The method was useful for samples of amino acids which liberated from 0.06 to 0.3 mgm. of carbon dioxide. The average deviation from the mean was somewhat less than 1% for samples liberating 0.2 mgm. of carbon dioxide. Use of the method for routine analysis of enzymatic digests resulted in substantial saving of time and material.


1987 ◽  
Vol 247 (1) ◽  
pp. 15-21 ◽  
Author(s):  
J Kellermann ◽  
C Thelen ◽  
F Lottspeich ◽  
A Henschen ◽  
R Vogel ◽  
...  

The arrangement of the disulphide bridges in human low-Mr kininogen has been elucidated. Low-Mr kininogen contains 18 half-cystine residues forming nine disulphide bridges. The first and the last half-cystine residues of the amino acid sequence form a disulphide loop which spans the heavy- and the light-chain portion of the kininogen molecule. The other 16 half-cystine residues are linked consecutively to form eight loops of 4-20 amino acids; these loops are lined up in the heavy-chain portion of the kininogen molecule. In this way, a particular pattern of disulphide loops is formed which seems to be of critical importance for the inhibitor function of human kininogen.


2015 ◽  
Vol 154 (3) ◽  
pp. 438-455 ◽  
Author(s):  
Q. S. ZUO ◽  
G. S. ZHOU ◽  
S. F. YANG ◽  
Y. YANG ◽  
L. R. WU ◽  
...  

SUMMARYCanola (Brassica napusL.) meal is widely used in animal feed as a protein source, and its quality relies on protein and amino acid content. However, little information is available regarding amino acid regulation in canola seed with nitrogen (N) application. The present study is aimed to evaluate the effect of N rate and genotype on canola seed amino acid concentrations under field conditions. A split-plot design comprising four N rates (0, 120, 240 and 360 kg N/ha) and three genotypes differing in seed protein content were used in 2010/11 and 2011/12. The results showed that increasing N rate decreased seed oil content linearly but increased seed protein content in all of the genotypes. The total amino acid concentration and absolute concentrations of individual amino acids in canola seed also improved significantly with the N rates in all of the genotypes. Regarding the proportions of amino acids, a group that included glutamic acid (Glu), proline (Pro) and arginine (Arg) dominated and occupied > 0·30 compared with other amino acids. The ratio of amino acids in this group increased by 8·3% with 360 kg N/ha compared with the control. However, the proportions of the other amino acids showed negative responses to the N rates. The results of regression analysis of the responses of individual amino acids to N rate indicated that Glu, Pro and Arg had a greater improvement potential with application of N fertilizer, as revealed by higher slopes in the linear equations compared with the other amino acids. Additionally, the concentrations of sulphur-containing amino acids, methionine and cysteine, were also a potential target for improving with N application because these are always deficient in major crops. In conclusion, N application cannot only improve seed protein content but also enhance deposition of amino acids such as Glu, Pro and Arg.


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