The Biosynthesis of Salicylic Acid in Potato Plants1

Spraying potato (Solanum tuberosum L.) leaves with arachidonic acid (AA) at 1500 μg mL−1 led to a rapid local synthesis of salicylic acid (SA) and accumulation of a SA conjugate, which was shown to be 2-O-β-glucopyranosylsalicylic acid. Radiolabeling studies with untreated leaves showed that SA was synthesized from phenylalanine and that both cinnamic and benzoic acid were intermediates in the biosynthesis pathway. Using radiolabeled phenylalanine as a precursor, the specific activity of SA was found to be lower when leaves were treated with AA than in control leaves. Similar results were obtained when leaves were fed with the labeled putative intermediates cinnamic acid and benzoic acid. Application of 2-aminoindan-2-phosphonic acid at 40 μm, an inhibitor of phenylalanine ammonia-lyase, prior to treatment with AA inhibited the local accumulation of SA. When the putative intermediates were applied to leaves in the presence of 2-aminoindan-2-phosphonic acid, about 40% of the expected accumulation of free SA was recovered, but the amount of the conjugate remained constant.

Posttranslational amino acid epimerization: enzyme-catalyzed isomerization of amino acid residues in peptide chains.

Since ribosomally mediated protein biosynthesis is confined to the L-amino acid pool, the presence of D-amino acids in peptides was considered for many years to be restricted to proteins of prokaryotic origin. Unicellular microorganisms have been responsible for the generation of a host of D-amino acid-containing peptide antibiotics (gramicidin, actinomycin, bacitracin, polymyxins). Recently, a series of mu and delta opioid receptor agonists [dermorphins and deltorphins] and neuroactive tetrapeptides containing a D-amino acid residue have been isolated from amphibian (frog) skin and mollusks. Amino acid sequences obtained from the cDNA libraries coincide with the observed dermorphin and deltorphin sequences, suggesting a stereospecific posttranslational amino acid isomerization of unknown mechanism. A cofactor-independent serine isomerase found in the venom of the Agelenopsis aperta spider provides the first major clue to explain how multicellular organisms are capable of incorporating single D-amino acid residues into these and other eukaryotic peptides. The enzyme is capable of isomerizing serine, cysteine, O-methylserine, and alanine residues in the middle of peptide chains, thereby providing a biochemical capability that, until now, had not been observed. Both D- and L-amino acid residues are susceptible to isomerization. The substrates share a common Leu-Xaa-Phe-Ala recognition site. Early in the reaction sequence, solvent-derived deuterium resides solely with the epimerized product (not substrate) in isomerizations carried out in 2H2O. Significant deuterium isotope effects are obtained in these reactions in addition to isomerizations of isotopically labeled substrates (2H at the epimerizeable serine alpha-carbon atom). The combined kinetic and structural data suggests a two-base mechanism in which abstraction of a proton from one face is concomitant with delivery from the opposite face by the conjugate acid of the second enzymic base.

Involvement of cytochrome P450 in oxime production in glucosinolate biosynthesis as demonstrated by an in vitro microsomal enzyme system isolated from jasmonic acid-induced seedlings of Sinapis alba L.

An in vitro enzyme system for the conversion of amino acid to oxime in the biosynthesis of glucosinolates has been established by the combined use of an improved isolation medium and jasmonic acid-induced etiolated seedlings of Sinapis alba L. An 8-fold induction of de novo biosynthesis of the L-tyrosine-derived p-hydroxybenzylglucosinolate was obtained in etiolated S. alba seedlings upon treatment with jasmonic acid. Formation of inhibitory glucosinolate degradation products upon tissue homogenization was prevented by inactivation of myrosinase by addition of 100 mM ascorbic acid to the isolation buffer. The biosynthetically active microsomal enzyme system converted L-tyrosine into p-hydroxyphenylacetaldoxime and the production of oxime was strictly dependent on NADPH. The Km and Vmax values of the enzyme system were 346 microM and 538 pmol per mg of protein per h, respectively. The nature of the enzyme catalyzing the conversion of amino acid to oxime in the biosynthesis of glucosinolates has been subject of much speculation. In the present paper, we demonstrate the involvement of cytochrome P450 by photoreversible inhibition by carbon monoxide. The inhibitory effect of numerous cytochrome P450 inhibitors confirms the involvement of cytochrome P450. This provides experimental documentation of similarity between the enzymes converting amino acids into the corresponding oximes in the biosynthesis of glucosinolates and cyanogenic glycosides.

Inhibitors of human heart chymase based on a peptide library.

We have synthesized two sets of noncleavable peptide-inhibitor libraries to map the S and S' subsites of human heart chymase. Human heart chymase is a chymotrypsin-like enzyme that converts angiotensin I to angiotensin II. The first library consists of peptides with 3-fluorobenzylpyruvamides in the P1 position. (Amino acid residues of substrates numbered P1, P2, etc., are toward the N-terminal direction, and P'1, P'2, etc., are toward the C-terminal direction from the scissile bond.) The P'1 and P'2 positions were varied to contain each one of the 20 naturally occurring amino acids and P'3 was kept constant as an arginine. The second library consists of peptides with phenylalanine keto-amides at P1, glycine in P'1, and benzyloxycarbonyl (Z)-isoleucine in P4. The P2 and P3 positions were varied to contain each of the naturally occurring amino acids, except for cysteine and methionine. The peptides of both libraries are attached to a solid support (pins). The peptides are evaluated by immersing the pins in a solution of the target enzyme and evaluating the amount of enzyme absorbed. The pins with the best inhibitors will absorb most enzyme. The libraries select the best and worst inhibitors within each group of peptides and provide an approximate ranking of the remaining peptides according to Ki. Through this library, we determined that Z-Ile-Glu-Pro-Phe-CO2Me and (F)-Phe-CO-Glu-Asp-ArgOMe should be the best inhibitors of chymase in this collection of peptide inhibitors. We synthesized the peptides and found Ki values were 1 nM and 1 microM, respectively. The corresponding Ki values for chymotrypsin were 10 nM and 100 microM. The use of libraries of inhibitors has advantages over the classical method of synthesis of potential inhibitors in solution: the libraries are reusable, the same libraries can be used with a variety of different serine proteases, and the method allows the screening of hundreds of compounds in short periods of time.

Overexpression of csk inhibits acid-induced activation of NHE-3.

Opossum kidney OKP cells express an apical membrane Na+/H+ antiporter that is encoded by NHE-3 (for Na+/H+ exchanger 3) and is similar in many respects to the renal proximal tubule apical membrane Na+/H+ antiporter. Chronic incubation of OKP cells in acid medium for 24 hr increases Na+/H(+)-antiporter activity and NHE-3 mRNA abundance. The increase in Na+/H(+)-antiporter activity was not prevented by H7, a protein kinase C/protein kinase A inhibitor, but was prevented by herbimycin A, a tyrosine kinase inhibitor. Incubation of cells in acid medium increased c-src activity, and this was inhibited by herbimycin A. To determine the role of the src family of nonreceptor protein-tyrosine kinases, Csk (for carboxyl-terminal src kinase), a physiologic inhibitor of these kinases, was overexpressed in OKP cells. In three clones overexpressing csk, acid-induced increases in Na+/H(+)-antiporter activity and NHE-3 mRNA abundance were inhibited. In these clones, inhibition of acid activation of Na+/H(+)-antiporter activity paralleled inhibition of acid activation of c-src. Neither herbimycin A nor overexpression of csk inhibited dexamethasone-induced increases in Na+/H(+)-antiporter activity. These studies show that decreases in pH activate c-src and that the src family nonreceptor protein-tyrosine kinases play a key role in acid activation of NHE-3.

Non-spectral interferences due to the presence of sulfuric acid in inductively coupled plasma mass spectrometry

Results of a systematic study concerning non-spectral interferences from sulfuric acid containing matrices on a large number of elements in inductively coupled plasma–mass spectrometry (ICP-MS) are presented in this work. The signals obtained with sulfuric acid solutions of different concentrations (up to 5% w w− 1) have been compared with the corresponding signals for a 1% w w− 1− nitric acid solution at different experimental conditions (i.e., sample uptake rates, nebulizer gas flows and r.f. powers). The signals observed for 128Te+, 78Se+ and 75As+ were significantly higher when using sulfuric acid matrices (up to 2.2-fold for 128Te+ and 78Se+ and 1.8-fold for 75As+ in the presence of 5 w w-1 sulfuric acid) for the whole range of experimental conditions tested. This is in agreement with previously reported observations. The signal for 31P+ is also higher (1.1-fold) in the presence of sulfuric acid. The signal enhancements for 128Te+, 78Se+, 75As+ and 31P+ are explained in relation to an increase in the analyte ion population as a result of charge transfer reactions involving S+ species in the plasma. Theoretical data suggest that Os, Sb, Pt, Ir, Zn and Hg could also be involved in sulfur-based charge transfer reactions, but no experimental evidence has been found. The presence of sulfuric acid gives rise to lower ion signals (about 10–20% lower) for the other nuclides tested, thus indicating the negative matrix effect caused by changes in the amount of analyte loading of the plasma. The elemental composition of a certified low-density polyethylene sample (ERM-EC681K) was determined by ICP-MS after two different sample digestion procedures, one of them including sulfuric acid. Element concentrations were in agreement with the certified values, irrespective of the acids used for the digestion. These results demonstrate that the use of matrix-matched standards allows the accurate determination of the tested elements in a sulfuric acid matrix.

Gakara region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4673-I (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Gakara. Printed in: 1994. Covers portions of Gakara region, Burundi. Sheet: 4673-I. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Bubanza region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4674-I (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Bubanza. Printed in: 1994. Covers portions of Bubanza region, Burundi. Sheet: 4674-I. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Bujumbura region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4674-II (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Bujumbura. Printed in: 1994. Covers portions of Bujumbura region, Burundi. Sheet: 4674-II. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Butahana region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4675-I (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Butahana. Printed in: 1994. Covers portions of Butahana region, Burundi. Sheet: 4675-I. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Ndora region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4675-II (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Ndora. Printed in: 1994. Covers portions of Ndora region, Burundi. Sheet: 4675-II. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Cibitoke region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4675-III (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Cibitoke. Printed in: 1994. Covers portions of Cibitoke region, Burundi. Sheet: 4675-III. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Mabayi region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4675-IV (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Mabayi. Printed in: 1994. Covers portions of Mabayi region, Burundi. Sheet: 4675-IV. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Mabanda region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4772-II (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Mabanda. Printed in: 1994. Covers portions of Mabanda region, Burundi. Sheet: 4772-II. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

Nyanza-Lac region, Burundi, 1994, Defense Mapping Agency (DMA) Series Z724, Sheet 4772-III (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Z724, Burundi, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Nyanza-Lac. Printed in: 1994. Covers portions of Nyanza-Lac region, Burundi. Sheet: 4772-III. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Burundi 1:50:000 Series Z724 maps are in English and French (legends also include Rundi). Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 20 meters, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.