Biosynthesis of phytosiderophores, mugineic acids, associated with methionine cycling.

The biosynthesis of 2'-deoxymugineic acid, a key phytosiderophore, was examined in association with the putative methionine recycling pathway in the roots of wheat using labeling experiments and structural analysis. Feeding with D-[1-13C]ribose did not result in 13C enrichment of 2'-deoxymugineic acid, while D-[2-13C]ribose resulted in 13C enrichment at the C-4", -1, -4' positions, and D-[5-13C]ribose did in C-1', -4, and -1" positions of 2'-deoxymugineic acid, respectively. Furthermore, two isotope-labeled intermediates of the methionine recycling pathway, 5-[5-2H2]methylthioribose and 2-[1-13C]keto-4-methylthiobutyric acid, were synthesized, and their incorporation into 2'-deoxymugineic acids was investigated.

A novel cDNA sequence encoding the precursor of the D-amino acid-containing neuropeptide fulicin and multiple alpha-amidated neuropeptides from Achatina fulica.

Fulicin (Phe-D-Asn-Glu-Phe-Val-NH2) is an endogenous neuropeptide containing a D-amino acid from ganglia of the African giant snail Achatina fulica. We have cloned a novel cDNA (1,995 nucleotides) encoding a fulicin precursor from the snail cerebral and subesophageal ganglia. The fulicin precursor protein (357 amino acids) contains one copy of fulicin and at least nine other putative alpha-amidated neuropeptides composed of four to six amino acid residues.

Characterization of crustacean hyperglycemic hormone from the crayfish (Procambarus clarkii): multiplicity of molecular forms by stereoinversion and diverse functions.

The complete amino acid sequence of two forms of crustacean hyperglycemic hormone (CHH) from the X-organ sinus gland complex of crayfish (Procambarus clarkii) has been determined. There are two variants of P. clarkii CHH (Prc-CHH), I and II, which can be separated by reversed-phase high-performance liquid chromatography (RP-HPLC).

Incorporation of Label from 13C-, 2H-, and 15N-Labeled Methionine Molecules during the Biosynthesis of 2[prime]-Deoxymugineic Acid in Roots of Wheat.

The biosynthetic pathway of 2[prime]-deoxymugineic acid, a key phytosiderophore, was investigated by feeding 13C-, 2H-, and 15N-labeled methionine, the first precursor, to the roots of hydroponically cultured wheat (Triticum aestivum L. cv Minori). The incorporation of label from each methionine species was observed during their conversion to 2[prime]-deoxymugineic acid, using 2H-, 15N-, and 13C-nuclear magnetic resonance (NMR).

Photosynthesis modulates the sign of phototaxis of wild-type Chlamydomonas reinhardtii. Effects of red background illumination and 3-(3',4'-dichlorophenyl)-1,1-dimethylurea.

We have found that the sign of phototaxis, i.e. the direction of phototactic migration either towards or away from the light source, of wild-type Chlamydomonas reinhardtii depends on its photosynthetic activity.

Two Related Biosynthetic Pathways of Mugineic Acids in Gramineous Plants.

The biosynthesis of mugineic acids was studied by feeding 2H- or 13C-labeled compounds to water-cultured roots in several gramineous plants. The fate of labeled compounds was monitored by using 2H- and 13C-nuclear magnetic resonance. On investigating the proton changes during biosynthesis by feeding D,L-[3,3,4,4-d4]-methionine (98.

WWamide-1, -2 and -3: novel neuromodulatory peptides isolated from ganglia of the African giant snail, Achatina fulica.

Three novel neuropeptides, isolated from ganglia of the African giant snail, Achatina fulica, were named WWamide-1, -2 and -3. These substances were biologically active heptapeptide amides with a Trp residue at both the N- and C-termini. WWamide-1, which displayed an inhibitory activity on a central neuron of the snail, exhibited peripherally modulatory effects on muscular contractions of not only the gut and other tissues of the snail but also certain tissues of other molluscs.

Two novel tachykinin-related neuropeptides in the echiuroid worm, Urechis unicinctus.

Two novel neuropeptides, urechistachykinin I (H-Leu-Arg-Gln-Ser-Gln-Phe-Val-Gly-Ser-Arg-NH2) and urechistachykinin II (H-Ala-Ala-Gly-Met-Gly-Phe-Phe-Gly-Ala-Arg-NH2), were isolated from the ventral nerve cords of the echiuroid worm, Urechis unicinctus. These peptides showed a contractile action on the inner circular body-wall muscle of the animal. Their amino acid sequences were found to be significantly homologous with those of the vertebrate and insect tachykinins.

Isolation of Antho-RFamide related peptides from the eyestalks of blue crab.

1. Two carboxyl-terminally amidated peptides (CP1 and CP2) were isolated from the blue crab (Callinectes sapidus) eyestalks by a method of carboxyl-terminal analysis. 2.

Diversion of the sign of phototaxis in a Chlamydomonas reinhardtii mutant incorporated with retinal and its analogs.

The blind mutant FN68 of the unicellular flagellate green alga Chlamydomonas reinhardtii is negatively phototactic in the presence of the native chromophore all-trans retinal. In contrast, analog chromophores such as a ring-acyclic retinal and those in which trans/cis isomerization about the C11 = C12 double bond was blocked induced predominantly positive phototaxis in the same strain under the same experimental conditions. These observations can be interpreted by assuming that the negative and the positive phototaxis is mediated distinctively by two rhodopsin species which differ in their affinities with the exogenous chromophores.

Structure and activity studies of pardaxin and analogues using model membranes of phosphatidylcholine.

Thirteen synthetic pardaxin analogues were assayed for their ability to interact with model membranes of phosphatidylcholine. The results suggested the following: An amphipathic alpha-helix from isoleucine-14 to leucine-26 is responsible for most of the membrane perturbing properties of pardaxin. A hydrophobic N-terminal region enhances the activity of the isoleucine-14 to leucine-26 alpha-helix by binding the pardaxin molecule to the lipid bilayer.

Sensitivity increase in the photophobic response of Halobacterium halobium reconstituted with retinal analogs: a novel interpretation for the fluence-response relationship and a kinetic modeling.

Phoborhodopsin (also called sensory rhodopsin II) is a photoreceptor protein which mediates photophobic responses of Halobacterium halobium to blue-green light. Under conditions where the synthesis of the chromophore retinal is inhibited, the photophobic system is reconstituted in vivo by incorporation of all-trans retinal or retinal analogs into the apoprotein of phoborhodopsin. Retinal analogs which retard the cyclic photoreaction kinetics of phoborhodopsin increase significantly the sensitivity of the photophobic response.

Diversity in steroidogenesis of symbiotic microorganisms from planthoppers.

Ergosta-5,7,24(28)-trien-3β-ol (trienol)6 was isolated from the intracellular symbiotes (symbiotic microorganisms) of the planthoppers,Nilaparvata lugens andLaodelphax striatellus. The steroidogenic end product of the symbiotes was found to depend on the environmental conditions, i.e.

Conformational feature of neuroactive domoic acid: X-ray structural comparison with isodomoic acid A and alpha-kainic acid.

As an aid for developing a new type of potent insecticide acting on the neuromuscular junction, conformational characteristics of domoic acid and isodomoic acid A, the naturally occurring glutamate agonists, were investigated by X-ray crystal analyses. Conformational comparison with a neuroactive alpha-kainic acid provides information concerning the stereochemical feature responsible for the biological activity.

Primary sequence confirmation of the beta-amyloid peptide from hereditary cerebral haemorrhage with amyloidosis (Dutch type) by collisional activation of the doubly charged ion.

High-energy collisional activation of doubly charged peptide ions allows direct sequential analysis of peptides over 4 kDa, including leucine/isoleucine determinations. This technique was used to verify the sequence of an intact beta-amyloid peptide associated with hereditary cerebral haemorrhage with amyloidosis Dutch type (4.2 kDa), which resists both enzymatic digestion and Edman sequencing.

NMR studies of amyloid beta-peptides: proton assignments, secondary structure, and mechanism of an alpha-helix----beta-sheet conversion for a homologous, 28-residue, N-terminal fragment.

Beta-peptide is a major component of amyloid deposits in Alzheimer's disease. We report here a proton nuclear magnetic resonance (NMR) spectroscopic investigation of a synthetic peptide that is homologous to residues 1-28 of beta-peptide [abbreviated as beta-(1-28)]. The beta-(1-28) peptide produces insoluble beta-pleated sheet structures in vitro, similar to the beta-pleated sheet structures of beta-peptide in amyloid deposits in vivo.

Solution conformations and aggregational properties of synthetic amyloid beta-peptides of Alzheimer's disease. Analysis of circular dichroism spectra.

The A4 or beta-peptide (39 to 43 amino acid residues) is the principal proteinaceous component of amyloid deposits in Alzheimer's disease. Using circular dichroism (c.d.

Covalent modification of Alzheimer's amyloid beta-peptide in formic acid solutions.

beta-peptide is a normal component of amyloid deposits in Alzheimer's disease. In aqueous solution, beta-peptide is extremely insoluble and rapidly aggregates forming oligomeric beta-sheet structures that eventually precipitate from solution. Presumably, this process is related to the production of amyloid deposits in Alzheimer's disease.

Tandem mass spectrometric analysis of peptides at the femtomole level.

In the sequential analysis of peptides by fast atom bombardment (FAB) tandem mass spectrometry (MS/MS), the principal obstacle to decreasing sample quantities was determined to be the signal-to-background ratio of the ionization/desorption process. By decreasing the background ion current, continuous-flow FAB allows complete analysis (both MS and MS/MS) to be performed on 25-75% less sample than required for a conventional FABMS/MS experiment alone. The combination of CF-FAB with array detection permitted sequential analysis of several peptides (900-2000 Da) at the 900 fmol to 5.

Photoisomerization of retinal at 13-ene is important for phototaxis of Chlamydomonas reinhardtii: simultaneous measurements of phototactic and photophobic responses.

A real-time automated method was developed for simultaneous measurements of phototactic orientation (phototaxis) and step-up photophobic response of flagellated microorganisms. Addition of all-trans retinal restored both photoresponses in a carotenoid-deficient mutant strain of Chlamydomonas reinhardtii in a dose-dependent manner. The phototactic orientation was biphasic with respect to both the light intensity and the concentration of retinal.

O-H stretching vibration in Fourier transform difference infrared spectra of bacteriorhodopsin.

FTIR difference spectroscopic studies of M intermediate and LA bacteriorhodopsin in the O-H stretching region show bands at 3671 and 3641 cm-1, respectively. The O-H stretching bands in this region may reflect protonation-deprotonation changes or environmental change in the tyrosine residues in bR.

Solution structures of beta peptide and its constituent fragments: relation to amyloid deposition.

The secondary structures in solution of the synthetic, naturally occurring, amyloid beta peptides, residues 1 to 42 [beta (1-42)] and beta (1-39), and related fragments, beta (1-28) and beta (29-42), have been studied by circular dichroism and two-dimensional nuclear magnetic resonance spectroscopy. In patients with Alzheimer's disease, extracellular amyloid plaque core is primarily composed of beta (1-42), whereas cerebrovascular amyloid contains the more soluble beta (1-39). In aqueous trifluoroethanol solution, the beta (1-28), beta (1-39), and beta (1-42) peptides adopt monomeric alpha-helical structures at both low and high pH, whereas at intermediate pH (4 to 7) an oligomeric beta structure (the probable structure in plaques) predominates.

Fourier transform infrared spectroscopic study on retinochrome and its primary photoproduct, lumiretinochrome.

Structural studies of retinochrome, and its photoproduct, lumiretinochrome, were done by Fourier transform infrared difference spectroscopy. The absorption bands in the carbonyl stretching region which shift in D2O show the changes in the protein part during the photoreaction. Strong absorption bands in the finger-print region show that the all-trans-retinal chromophore in retinochrome isomerizes to the 11-cis-retinal chromophore in lumiretinochrome upon illumination with yellow-green light at 83K.

Physiological role of 3-hydroxykynurenine and xanthurenic acid upon crustacean molting.

Studies with crabs (Charybdis japonica) and crayfish (Procambarus clarkii) revealed that the tryptophan metabolites, 3-hydroxy-L-kynurenine (3-OH-K) and xanthurenic acid (XA), common secretory products of the X-organ-sinus gland complex of eyestalks from several decapods, regulated the molting of crustaceans in species-nonspecific fashion. Injection of 3-OH-K to the eyestalk-ablated crayfish delayed the onset of the first molt and lengthened the interval between the first and second molts. These lines of evidence were in accord with previous accounts of the so-called "molt inhibiting hormone" (MIH) effect.