A novel monoantennary complex-type sugar chain found in octopus rhodopsin: occurrence of the Gal{beta}1->4Fuc group linked to the proximal N-acetylglucosaniine residue of the trimannosyl core

The N-linked sugar chains were liberated as oligosaccha-ridesfrom octopus rhodopsin by hydrazinolysis. Most of the oligosaccharideswere neutral, and separated into two major components by columnchromatography using immobilized lectins and Bio-Gel P-4. Structuralanalysis of the one major component by sequential exoglycosidasedigestion, chemical fragmentation in combination with meth-ylationanalysis revealed that it is a nonasaccharide; Man16(Gaiβ13GlcNAcβ12Man13)Manβ14GlcNAcβ14(Galβ14Fuc16)GlcNAcThis structure is quite unique in that a novel galactosylatedfucose residue is attached to the reducing terminal N-acetyl-glucosamineresidue. galactosylated Fuc N-linked sugar chain novel structure octopus rhodopsin     

Further characterization of the binding properties of a GalNAc specific lectin from Codium fragile subspecies tomentosoides

Previous study on the binding properties of a lectin isolatedfrom Codium fragile subspecies tomentosoides (CFT) indicatesthat this lectin recognizes the GalNAc1 sequence at both reducingand nonreducing ends. In this study, the carbohydrate specificityof CFT was further characterized by quantitative precipitin(QPA) and inhibition of lectin-enzyme binding assays. Of theglycoforms tested for QPA, all asialo-GalNAc1 containing glyco-proteinsreacted well with the lectin. Asialo hamster and ovine submandibularglycoproteins, which contain almost exclusively Tn (GalNAclSer/Thr)residues as carbohydrate side chains, and Streptococcus typeC polysaccharide completely precipitated the lectin added, whilethe GalNAcβcontaining Tamm-Horsfall Sd(a⁺) glycopro-teinand its asialo product were inactive. Among the oligo-saccharidestested for inhibiting lectin-glycoprotein interaction, GalNAc13GalNAcβ13Gal14Galβ14GIc(Fp)and Galβ13GalNAc1benzyl (T) were the best, and about 125-foldmore active than GalNAc They were about 3.3, 6.6, and 43 timesmore active than Tn containing glycopeptides, GalNAc13(LFuc12)Gal(A_h) and Galβ13GalNAc(T), respectively. From the presentand previous results, it is concluded that the combining siteof CFT is probably of a groove type that recognizes from GalNAclto pentasaccharide(Fp). The carbohydrate specificity of thislectin can be constructed and summarized in decreasing orderby lectin determinants as follows: Fp and T > Tn cluster> A_h >>I/II. carbohydrate specificities Codium fragile tomentosoides glycoprotein binding lectins     

Incorporation of some possible radioactive intermediates into chlorogenic acid in sliced sweet potato tissue

t-Cinnamic acid-2-¹⁴C, p-coumaric acid-2-¹⁴C and caffeic acid-2-¹⁴Cwere administered to discs of sweet potato roots and incorporationof each radioactive compound into chlorogenic acid was compared.The data suggest that chlorogenic acid is synthesized througheither or both of two major pathways, phenylalanine t-cinnamate t-cinnamoyl derivative p-coumaroyl derivative chlorogenicacid and phenylalanine t-cinnamate p-coumarate p-coumaroylderivative chlorogenic acid. ¹Part 75 of the phytopathological chemistry of sweet potatowith black rot and injury. ²Present address : Department of Biology, Tokyo MetropolitanUniversity, Setagaya-ku, Tokyo. (Received December 23, 1968; )     

Effects of auxin on the structure of hemicelluloses of Avene coleoptiles

Avena coleoptile hemicelluloses were fractionated into water-solublehemicelluloses I and IIB and water-insoluble hemicellulose IIA.These hemicelluloses were then subjected to glycosidic linkageanalysis by methylation technique, which revealed that hemicelluloseI was predominantly composed of arabinoxylans and ß-(l4)glucans and hemicellulose IIB was composed of arabinoxylans,ß-(l4) : (l3)-mixed linked glucans, ß-(l4)-glucansand xyloglucans. Hemicellulose IIA was mainly composed of xyloglucansand probably ß-(l4)-glucans. Methylation analysisof hemicelluloses extracted from Avena coleoptile segments treatedwith auxin in the presence of mannitol (0.15 M) indicated thatauxin apparently had no effect on the structure of arabinoxylanand caused a specific decrease in the amount of ß-(l4): (l3)-mixed linked glucan. (Received November 19, 1979; )     

Effects of illumination on absorption peak shifts in spectra of intact etiolated cotyledons of Pharbitis nil I. Existence of two kinds of shift patterns

Two patterns were found in the shifts of absorption peaks inspectra of intact etiolated Pharbitis cotyledons illuminatedat room temperature. One was a well-known pattern, P649C678C683C672,called the "high-intensity illumination pattern" in this study.The other, called the "low-intensity illumination pattern,"was P649C672. (Received June 16, 1976; )     

Comparative Biochemistry of the Pineal Gland

The pineal gland is biochemically very active. In mammals, ithas the unique capacity to synthesize the hormone melatonin(N-acetyl-5-methoxy-tryptamine). Although the synthesis of melatoninis confined mainly to the pineal gland of all vertebrates, theeyes and brains of amphibians and fish also can form melatonin.Melatonin is synthesized in the pineal as follows: tryptophan 5-hydroxy-tryptophan serotonin N-acetylserotonin melatonin.The final step is catalyzed by the enzyme, hydroxyindole-O-methyltransferase (HIOMT), which is highly localized in the pinealof all vertebrate species examined. The activity of HIOMT ischanged when animals are kept in constant darkness or light.In rats, highest HIOMT activity is present in constant darkness,while the reverse occurs in avian species. In mammals, informationabout lighting reaches the pineal via the retina inferior accessoryoptic tract preganglionic sympathetic fibers superior cervicalganglia postganglionic fibers pineal parenchymal cells. Lightingmessages reach the hen's pineal via a nonretinal pathway. Studieswith tissue culture indicate that noradrenaline liberated fromsympathetic nerves stimulates synthesis of melatonin. Thereare circadian rhythms in pineal serotonin content which areendogenous and abolished by removal of superior cervical gangliaor by decentralization. There is also a 24-hour rhythm in pinealnoradrenaline. This rhythm is exogenous and is abolished byblinding or cutting the inferior accessory optic tract.     

Xyloglucan and r{beta}-D-Glucan in Cell Walls of Rice Seedlings

Cell walls of 4-day old rice seedlings were extracted successivelywith ammonium oxalate-oxalic acid, 4% KOH and 24% KOH. A rß-D-glucanpreparation and a xyloglucan preparation were isolated fromthe 4% KOH extract and 24% KOH extract, respectively. Methylationanalysis and enzymic degradation studies of the polysaccharidesshowed that the former was built up predominantly of repeating-oligosaccharideunits of 3-O-rß-cellobiosyl-D-glucose and 3-O-rß-cellotriosyl-D-glucosein a molar ratio of 2.6 : 1.0, and the latter was of repeating-oligosaccharideunits of -D-xylosyl-(16)-rß-D-glucosyl-(14)-[-D-xylosyl-(16)]-rß-D-glucosyl-(14)-D-glucose,-D-xylosyl-(16)-rß-D-glucosyl-(14)-D-glucose and cellobiose. ¹ Present address: Department of Botany, Iowa State University,Ames, Iowa 50011, U.S.A. (Received August 29, 1981; Accepted January 12, 1982)     

AGMATINE AND N-CARBAMYLPUTRESCINE AS INTERMEDIATES IN THE FORMATION OF NICOTINE BY TOBACCO PLANTS

Agmatine-G-³H and N-carbamylputrescine-l,4-¹⁴C were effectivelyincorporated into the nicotine of tobacco plants. This resultmay indicate a route that the pyrrolidine ring of nicotine isformed from putrescine by the following pathway: arginineagmatineN-carbamylputrescineputrescinepyrrolidinering. (Received February 7, 1966; )     

Separation of partially desialylated branched oligosaccharide isomers containing {alpha}(2<-3)- and {alpha}(2<-6)-linked Neu5Ac

The following two tri-sialylated triantennary oligosaccharides,which differ only in the linkage of the Neu5Ac to the uppermostbranch were, individually, partially desialylated to produceall possible di- and mono-sialylated isomers. A tetra-sialylated triantennary isomer, which contained an (26)-linkedNeu5Ac to the GlcNAc on branch III, was also converted to allpossible trisialylated isomers by mild acid hydrolysis as previouslydescribed (Roher et al., Anal Biochem., 212, 7–16, 1993).The resulting branch isomers were separated using high-pH anion-exchangechromatography (HPAEC). Structures were assigned to peak fractionson the basis of the previously described effect of (26)- and(23)-linked Neu5Ac on the elution order of branched lactosamine-typeoligosaccharides (Townsend et al., Anal Biochem., 182, 1–8,1989). No differences in the acid lability of the Neu5Ac linkageto either Gal ((23) or (26)) or GlcNAc ((26)) were observed.Our studies show that chemical desialylation and HPAEC is auseful approach to prepare and identify all possible sialylatedbranch isomers and should prove useful for defining the branchspecificity of sialyltransferases and sialidases. high-pH anion-exchange chromatography pulsed electrochemical detection sialidases sialylated oligosaccharides sialyltransferases     

Interrelations between Phyllotaxis, Leaf Development and the Primary-secondary Vascular Transition in Populus deltoides

The procambial system of Populus deltoides Bartr. ex Marsh.plants progressed from phyllotaxy in the cotyledon stage throughthe phyllotactic orders 3/2;5/13. The nodal position at whicheach of these phyllotactic transitions occured was determinedby anatomical analyses; they were found to be remarkably consistentin a large population of young plants. The data were used todiagrammatically reconstruct the procambial system of a typical16 leaf plant. Because all plant parts grew continuously anduninterruptedly, it was not possible to verify the positionsof the phyllotactic transitions by morphological criteria. However,several measured parameters (the number and lengths of primordiawithin the terminal bud, the plastochron interval, and the numberof leaf traces with birefringent xylem elcments) attained constantvalues following establishment of the 5/13 phyllotaxy, suggestingthis to be the stable phyllotactic order for the species. Althoughbud size continued to increase in plants exhibiting 5/13 phyllotaxy,it could be accounted for by the increased number and size ofbasipetal subsidiary bundles in the procambial leaf traces.It was suggested that these phyllotactic transitions in theprocambial system are programmed in the plant to occur at ratherspecific stages of ontogeny. The process is mediated by theolder leaves and it is therefore modified by plant vigour. Locationof the primary-secondary vascular transition zone was also relatedto the order of phyllotaxy. It advanced acropetally in the stemin close association with leaf maturation, but this associationwas further influenced by plant vigour. Populus deltoides Bartr. ex Marsh., cottonwood, vascular anatomy, phyllotaxis, leaf growth, xylem     

The Structure and Functions of Xyloglucan

Xyloglucan is a polysaccharide found in the primary cell wallsof all higher plants examined. Its cellulose-like backbone,which is about 0.15 to 1.5 µm long, consists of 300 to3 000 ß-(14)-linked D-glucopyranose residues. About60–75% (or, in grasses, about 30–40%) of the glucoseresidues have side-chains attached to position 6. The majorside-chains are: D-xylopyranosyl--1 -, D-galactopyranosyl-ß-(12)-D-xylopyranosyl--I , L-arabinofuranosyl-(1 -2)-D-xylopyranosyl--1-, and (except in grasses) L.-fucopyranosyl--(1 -2)-D-galactopyranosyl-ß-(1-2)-D-xylopyranosyl--1-. There is some regularity in the distribution of these side-chainsalong the backbone. Xyloglucan plays two very different r?les in the control ofcell growth: (a) as a major building material of the wall [concentrationof xyloglucan in the wall in vivo 10% (w/v)] it probably directlydictates wall extensibility and, therefore, the rate of cellexpansion and (b) it can be broken down to a fucose-containingoligosaccharide which [at a concentration of 0.0000001% (w/v)]exerts a hormone-like anti-auxin effect on growth. In addition,xyloglucan lacking fucose is used by certain dicotyledonousseeds as a food reserve which is mobilized after germination.Xyloglucan is, therefore, the subject of considerable currentinterest in several apparently disparate areas of botany. Key words: Xyloglucan, ‘oligosaccharin’, hemicellulose, auxin, anti-auxin, growth, cell walls, reserve carbohydrate     

Symmetry and Cell Division in Raphid Diatoms

The development and final morphology of the valve in raphe-bearingdiatoms exhibit a cryptic lateral polarity, and hence two typesof frustule can be distinguished. In the cis type both valveshave the same orientation; in the trans type they have oppositeorientations. Examination of a variety of taxa suggests thatin all dividing raphid diatoms, both new valves have the sameorientation and so only three types of division are possible:cis; cis + cis, cis trans + trans, trans; cis+trans. The possessionof different combinations of these explains the observed ratiosof cis: trans in different taxa, viz all cis; 1: 2 cis: trans;and roughly 1: 1 cis: trans. The implications of the resultsfor diatom systematics are examined, with special referenceto Navicula Bory. Diatom systematic, diatom valve morphogenesis, cell symmetry, raphe structure     

Chlorophyll Formation in the YG-6 Mutant of Chlorella regularis: Spectral Characterization of Protochlorophyllide Phototransformation

Dark-grown YG-6 mutant cells of Chlorella regularis accumulateat least two forms of phototransformable protochlorophyllide(Pchlide) with in vivo absorption maxima at 634 nm (Pchlide634) and 650 nm (Pchlide 650). Difference spectrophotometricanalyses and the action spectra showed that Pchlide 634 is firsttransformed into the 648 nm form and then phototransformed intochlorophyllide (Chlide) 672 nm. Pchlide 650 is phototransformedinto Chlide 685 which then shifts towards short wavelength-formingChlide 667 in the subsequent dark stage (Shibata shift). Pchlide650 is regenerated at the expense of photoinactive Pchlide 632.In washed cells after the phototransformation, the Shibata shiftwas accelerated. Freezing/thawing treatment in the dark causedconversion of phototransformable Pchlide 650 into photoinactivePchlide 633, but phototransformation activity of Pchlide 634still partly remained. These results suggest that in the final step of light-dependentchlorophyll formation in the YG-6 mutant of C. regularis, twosequentially and functionally separate routes are present: (1) Pchlide 634 Pchlide 648 Chlide 672 Chlorophyll a. (2) Pchlide 650 Chlide 685 Chlide 667 Chlorophyll a. (Received June 4, 1983; Accepted November 11, 1983)     

Mycobacterial arabinan biosynthesis: the use of synthetic arabinoside acceptors in the development of an arabinosyl transfer assay

Information on the biosynthesis of the D-arabinans of the cellwall of Mycobacterium tuberculosis is rapidly emerging, withthe promise of new targets for drug development against tuberculosis.Accordingly, arabinosyl transferase assays were developed utilizingsynthesized [1–¹⁴C]-β-D-arabinofuranosyl-1-monophosphoryldecaprenolas donor and a variety of O- and S-alkyl arabinosides as acceptors.These were: -D-Araf-(15)--D-Araf-O- and -S-alkyl di-arabinosidesand -D-Araf-(15)--D-Araf-(15)--D-Araf-O- and -S-alkyl triarabinosides.Whereas the O- and S-alkyl monosaccharide acceptors were inactive,the O- and S-alkyl disaccharide and the O- and S-alkyl trisaccharideacceptors (<C12) possessed considerable acceptor activity,and the trisaccharide acceptors were more potent than the correspondingdisaccharides. The O-alkyl disaccharide acceptors with a C8alkyl chain were more active than those containing the C6 orC10 analogs. Chemical analysis of the enzymatically synthesizedproducts of the reactions demonstrated that β-D-arabinofuranosyl-1-monophosphoryldecaprenolwas an effective donor for two of the three potential arabinosyltransferases: β-D-arabinofuranosyl-1-monophosphoryldecaprenol:arabinan (15) arabinosyl transferase and β-D-arabinofuranosyl-1-monophosphoryl-decaprenol:arabinan β(12) arabinosyl transferase. The β(12) arabinosyltransferase activity was more in evidence in the presence ofthe O-alkyl disaccharide acceptor, whereas both transferaseswere about equivalent in the presence of the S-alkyl trisaccharideacceptor. The tuberculosis drug, ethambutol, a known mycobacterialarabinosyl transferase inhibitor, was inactive within thesearabinosyl transferase/acceptor based assay systems, supportingother evidence that a third activity, responsible for the formationof 13 linkage, is the drug target. acceptor arabinan biosynthesis glycosyltrans-ferase assay mycobacteria     

Metabolism of Choline Chloride and Its Analogs in Wheat Seedlings

The incorporation rate of choline chloride and allylcholinebromide into wheat protoplasts were rapid compared with theincorporation rate of benzylcholine bromide. Choline chloridewas metabolized via two pathways: choline betaine and choline phosphorylcholine phos-phatidylcholine. Allylcholine bromidewas metabolized via only one pathway: allylcholine phosphorylallylcholine phosphatidylallylcholine, and benzylcholine bromide was notmetabolized at all. These results suggest that the stimulationof photosynthesis (Hyeon et al. 1988) by these compounds iscaused directly by these choline analogs and not by their metabolites. (Received June 29, 1989; Accepted October 20, 1989)     

Long chain fatty acid synthesis in developing castor bean seeds II. Operation of the path from sucrose to long chain fatty acids in a subcellular particulate system

Particles spun down at 10,000 ? g from developing castor beanseeds were capable of synthesizing LFAs from sucrose, a physiologicalprecursor transferred from leaves as a photosynthetic product.Tracer experiments, in combination with inhibitor effects, intermediatedilutions and cofactor requirements, indicated the operationof the following path: sucroseUDPGG-1-PG-6-PGAPpyruvateacetyl-CoAmalonyl-CoALFA.The whole path appears to be associated with 10,000 ? g particles,because repeated washings were unsuccessful in dissociatinga partial path from the particles, depsite of disorganizingthe structure of the particles. Based on the occurrence of freehexose(s) and the utilization of UDPG similar to that of sucroseor G-1-P in this reaction, it is probable that hexose phosphateis formed from sucrose via UDPG and fructose, although the conversionof sucrose to hexose phosphates via glucose and furctose isnot ruled out. Inhibitor experiments showed that ATP is self-supportingover the whole path, the ATP formed in the glycolytic path beingconsumed in a acetyl-CoA carboxylation step. Since oxidizedpyridine nucleotides are as available as reduced ones for LFAsynthesis, they seem to shuttle between the reduction in theconversion of sucrose to acetyl-CoA and the oxidation in LFAsynthesis from acetyl-CoA. From the pattern of the LFAs synthesized,NAD⁺ is available for the synthesis of saturated LFAs (18: 0,16: 0). whereas NADP⁺ is available for that of unsaturated LFAs(18: 1, 16: 1). (Received July 23, 1973; )     

Human Lewis {alpha}1->3/4fucosyltransferase: specificity of fucose transfer to GlcNAc{beta}1->3Gal{beta}1->4Glc{beta}1->1Cer (LcOse3Cer)

Biosynthesis of the Le^x series of carbohydrate antigens proceedsby fucose transfer in 13-linkage to the penultimate GlcNAc residueof a neolacto-series oligosaccharide acceptor, a reaction catalysedby multiple enzymes expressed in human tissues. Particularlybroad acceptor specificity, including the ability to catalysefucose transfer to both lacto- and neolacto-series acceptorsas well as the precursor Lc₃ structure (where Lc₃ lactotriaosylceramide,is GlcNAcß13Galß14Glcß1Cer), existsfor one human fucosyltransferase form, the Lewis 13/4fucosyltransferase(FucT-III). To determine if fucose transfer to Lc₃may representan alternate early step in Le^xor Le^a antigen biosynthesis withthis enzyme, the chemical structure of the fucosylated Lc₃ reactionproduct formed by the Lewis 13/4fucosyltransferase from Colo205 cells has been defined. Transfer of [¹⁴C]fucose to Lc₃ yieldeda labelled product migrating as a tetrasaccharide on thin layerchromatography plates. This product remained an acceptor forboth ß13- and ß14-galactosyl transfer onthe terminal GlcNAc residue. The product was degraded to a fucosylatedtrisaccharide derivative by bovine kidney ß-N-acetylglucosaminidase.Fast atom bombardment mass spectrometry and methylation analysisconfirmed that the product was composed exclusively of the followingstructure containing a fucose linked to the 3-position of theinternal Glc residue: GlcNAcß13Galß14Glcß11Cer Such a structure does not represent an intermdiate in Le^xorLe^a antigen biosynthesis. Thus, the evidence suggests that Le^xor Le^a antigen synthesis results exclusively from fucosylationof complete core chains. fucosyltransferase lacto-series LcOse₃Cer Lewis antigen transfer specificity     

Flexibility in the donor substrate specificity of {beta} 1,4-galactosyltransferase: application in the synthesis of complex carbohydrates

Biosynthetically, bovine N-acetylglucosainine ß 1,4-galacto-syltransferase(GalT) catalyses the transfer of galactosyl residues from UDP-Galto the 4-position of GlcNAc units, resulting in the productionof N-acetyllactosamine sequences. UDP-Glc and UDP-GalNAc werealso found to act as donors for this enzyme, allowing the preparationof ßGlc(14)-ßGlcNAc and ßGalNAc(14)ßGlcNActerminating structures on the milligram scale. GalT could thusbe used to add ßGalNAc to ßGlcNAc(12)Manterminating structures, converting them to the ßGalNAc(14)ßGlcNAc(12)Mansequences found on glycoprotein hormones. GalT did not transferGlcNAc residues from UDP-GlcNAc, but it could utilize UDP-GlcNH₂as a donor. Synthesis of ßGlcNAc(14)ßGlcNAcsequences could therefore be accomplished by transfer of GlcNH₂from its UDP derivative, followed by N-acetylation of the productamino-disaccharide using acetic anhydride in methanol. The productsof the enzymatic reactions were characterized by ¹H-NMR-spectroscopyand fast-atom bombardment mass spectrometry. This work expandsthe scope of the combined chemical-enzymatic synthesis of complexcarbohydrates, using glycosyltrans-ferases, to the productionof oligosaccharides different from those for which these enzymeswere designed. These unnatural reactions should find applicationin glycoprotein and glycolipid remodelling. galactosyltransferase chemica1-enzymatic synthesis of oligosaccharides oligosaccharide analogues sugar-nucleotide analogues carbohydrate remodelling     

Wilting of Leaves in Vicia faba L.

Wilting of leaves of Vicia faba L., which occurs when the pressurepotential (_p) is zero, and the leaf-water potential () at wiltingboth depend entirely upon the solute potential at incipientplasmolysis (_so) and not on soil-water status. Wilting in V.faba is acropetal; this is consistent with the hypothesis thatthere is a gradient of decreasing _so up the plant and that wateris transferred from the lower to the upper leaves, hasteningthe overall water loss from the lower leaves to the point when_p is zero. The gradient in _so up the plant is of the order of3–8 bar. It is proposed that wilting when _p>0 (i.e. > _so) shouldbe ‘apparent wilting’ and that when _p0 (i.e. _so),‘true wilting’.     

Rhizoid differentiation in Spirogyra I. Basic features of rhizoid formation

Several types of rhizoids occurring in the process of differentiationin Spirogyra sp. are described and their interrelation was elucidated.There are two differentiation sequences: P_pPRh_ros or PpPRh_rodRh_ros(for explanation of abbreviations see p. 533), although undersome conditions the sequences ceased halfway through. The initiationtime for rhizoid formation had no relation to the cell cyclestage. The difference in growth patterns between the rhizoidand ordinary filament cells was demonstrated with Calcofluor-stainingand centrifugation. The optimum temperature and pH of the culture medium for rhizoiddifferentiation were 20?C and pH 7, respectively. A contactstimulus was not necessary for induction. Of the several environmental factors examined, light was themost important, for rhizoid formation, since a rhizoid was inducedonly when light was given after cutting the filament. (Received December 14, 1972; )