Cloning, expression, purification, and characterization of the acid alpha-mannosidase from Trypanosoma cruzi.

Abstract:

:The acid alpha-mannosidase of Trypanosoma cruzi is a broad-specificity hydrolase involved in the catabolism of glycoconjugates, presumably in the digestive vacuole. We have cloned the alpha-mannosidase gene from a T.cruzi epimastigote genomic library. The alpha-mannosidase gene was determined to be single copy by Southern analysis, and similar sequences were not detected in genomic digests of either Trypanosoma brucei or Leishmania donovani. The coding region was subcloned into the Pichia pastoris expression vector pPICZ, and alpha-mannosidase activity was detected in the medium of induced cultures. The recombinant alpha-mannosidase demonstrated a pH optimum, inhibition by swainsonine, Km, and substrate specificity consistent with the characteristics of the alpha-mannosidase previously purified from T.cruzi epimastigotes. The recombinant enzyme was purified 103-fold from the culture medium of Pichia pastoris and had a native molecular mass of 359 kDa by gel filtration. A combination of SDS-PAGE, deglycosylation with endo H, and NH2-terminal sequencing indicates that the enzyme is originally synthesized as a homodimeric polypeptide that is subsequently cleaved to form a heterotetramer composed of 57 and 46 kDa subunits. A polyclonal antibody raised to the recombinant enzyme was shown to immunoprecipitate the alpha-mannosidase from T.cruzi cell extracts and will be used in future immunolocalization studies.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Vandersall-Nairn AS,Merkle RK,O'Brien K,Oeltmann TN,Moremen KW

doi

10.1093/glycob/8.12.1183

subject

Has Abstract

pub_date

1998-12-01 00:00:00

pages

1183-94

issue

12

eissn

0959-6658

issn

1460-2423

pii

cwb125

journal_volume

8

pub_type

杂志文章
  • Differential distribution of major gangliosides in rat central nervous system detected by specific monoclonal antibodies.

    abstract::We investigated the localization of major gangliosides in adult rat brain by an immunofluorescence technique with mouse monoclonal antibodies (MAbs). Five MAbs (GMB16, GMR17, GGR12, GMR5 and GMR13) that specifically recognize gangliosides GM1, GD1a, GD1b, GT1b and GQ1b, respectively, were used. We have found that ther...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/3.2.137

    authors: Kotani M,Kawashima I,Ozawa H,Terashima T,Tai T

    更新日期:1993-04-01 00:00:00

  • Sweating the small stuff: Glycoproteins in human sweat and their unexplored potential for microbial adhesion.

    abstract::There is increasing evidence that secretory fluids such as tears, saliva and milk play an important role in protecting the human body from infection via a washing mechanism involving glycan-mediated adhesion of potential pathogens to secretory glycoproteins. Interaction of sweat with bacteria is well established as th...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cwv102

    authors: Peterson RA,Gueniche A,Adam de Beaumais S,Breton L,Dalko-Csiba M,Packer NH

    更新日期:2016-03-01 00:00:00

  • Human sialidase NEU4 long and short are extrinsic proteins bound to outer mitochondrial membrane and the endoplasmic reticulum, respectively.

    abstract::Sialidases are widely distributed glycohydrolytic enzymes removing sialic acid residues from glycoconjugates. In mammals, several sialidases with different subcellular localizations and biochemical features have been described. NEU4, the most recently identified member of the human sialidase family, is found in two fo...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp156

    authors: Bigi A,Morosi L,Pozzi C,Forcella M,Tettamanti G,Venerando B,Monti E,Fusi P

    更新日期:2010-02-01 00:00:00

  • Induction of sialyl-Lex expression by herpes simplex virus type 1 is dependent on viral immediate early RNA-activated transcription of host fucosyltransferase genes.

    abstract::We have previously shown that varicella-zoster virus (VZV) and cytomegalovirus (CMV) infection of diploid human fibroblasts (HEL) results in neo-expression of Lewis antigens sialyl Lewis x (sLe(x)) and Lewis y (Le(y)), respectively, after transcriptional activation of different combinations of dormant human fucosyltra...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp057

    authors: Nyström K,Nordén R,Muylaert I,Elias P,Larson G,Olofsson S

    更新日期:2009-08-01 00:00:00

  • Inhibition of glycoprotein processing by L-fructose and L-xylulose.

    abstract::A number of unusual and rare carbohydrates were tested as potential inhibitors of various glycosidases, as well as inhibitors of N-linked oligosaccharide processing. The best inhibitors of several arylglycosidases and of glucosidase I were L-xylulose and L-fructose. Both of these sugars showed some inhibitory activity...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/6.8.795

    authors: Muniruzzaman S,Pan YT,Zeng Y,Atkins B,Izumori K,Elbein AD

    更新日期:1996-12-01 00:00:00

  • Elucidation of the O-antigen structure of Escherichia coli O63.

    abstract::The structure of the O-antigen polysaccharide (PS) from the Shiga-toxin producing Escherichia coli O63 has been elucidated using a combination of bioinformatics, component analyses and NMR spectroscopy. The O-antigen is comprised of tetrasaccharide repeating units with the following structure: →2)-β-d-Quip3N(d-allo-Th...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwy098

    authors: Ståhle J,Fontana C,Weintraub A,Widmalm G

    更新日期:2019-02-01 00:00:00

  • Apiose: one of nature's witty games.

    abstract::Apiose is a unique branched-chain pentose found principally in plants. It is a key component of structurally complex cell wall polysaccharides, as well as being present in a large number of naturally occurring secondary metabolites. This review provides a comprehensive overview of the current state of knowledge on the...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cww012

    authors: Pičmanová M,Møller BL

    更新日期:2016-05-01 00:00:00

  • Purification and cDNA cloning of Xenopus liver galectins and their expression.

    abstract::We have characterized galectin family proteins in adult tissues of Xenopus laevis and purified 14-kDa and 36-kDa proteins from the liver. The liver galectins showed comparable hemagglutination activities to those of mammalian galectins. Furthermore, we isolated five galectin cDNAs from a Xenopus liver library. These c...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/12.3.163

    authors: Shoji H,Nishi N,Hirashima M,Nakamura T

    更新日期:2002-03-01 00:00:00

  • Tn and STn are members of a family of carbohydrate tumor antigens that possess carbohydrate-carbohydrate interactions.

    abstract::The mucin-type O-glycome in cancer aberrantly expresses the truncated glycans Tn (GalNAcα1-Ser/Thr) and STn (Neu5Acα2,6GalNAcα1-Ser/Thr). However, the role of Tn and STn in cancer and other diseases is not well understood. Our recent discovery of the self-binding properties (carbohydrate-carbohydrate interactions, CCI...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cwy032

    authors: Sletmoen M,Gerken TA,Stokke BT,Burchell J,Brewer CF

    更新日期:2018-07-01 00:00:00

  • Production and characterization of a monoclonal antibody able to discriminate galectin-1 from galectin-2 and galectin-3.

    abstract::Antisera raised against galectin-1 exhibit crossreactivities with other galectins or related molecules. In order to overcome this problem, a monoclonal antibody to human brain galectin-1 was obtained by selecting clones without reactivity toward galectin-3. This mAb specifically bound galectin-1 of various animal orig...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/8.5.425

    authors: Cornillot JD,Pontet M,Dupuy C,Chadli A,Caron M,Joubert-Caron R,Bourin P,Bladier D

    更新日期:1998-05-01 00:00:00

  • Addition of beta1-6 GlcNAc branching to the oligosaccharide attached to Asn 772 in the serine protease domain of matriptase plays a pivotal role in its stability and resistance against trypsin.

    abstract::beta1-6 GlcNAc branching, a product of N-acetylglucosaminyltransferase V (GnT-V), is a key structure that is associated with malignant transformations and cancer metastasis. Although a number of reports concerning tumor metastasis-related glycoproteins that contain beta1-6 GlcNAc branching have appeared, the precise f...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh013

    authors: Ihara S,Miyoshi E,Nakahara S,Sakiyama H,Ihara H,Akinaga A,Honke K,Dickson RB,Lin CY,Taniguchi N

    更新日期:2004-02-01 00:00:00

  • Alpha1,4GlcNAc-capped mucin-type O-glycan inhibits cholesterol alpha-glucosyltransferase from Helicobacter pylori and suppresses H. pylori growth.

    abstract::Helicobacter pylori infects over half of the world's population and is thought to be a leading cause of gastric ulcer, gastric carcinoma, and gastric malignant lymphoma of mucosa-associated lymphoid tissue type. Previously, we reported that a gland mucin (MUC6) present in the lower portion of the gastric mucosa contai...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwn037

    authors: Lee H,Wang P,Hoshino H,Ito Y,Kobayashi M,Nakayama J,Seeberger PH,Fukuda M

    更新日期:2008-07-01 00:00:00

  • Cloning and characterization of a sialidase from the murine histocompatibility-2 complex: low levels of mRNA and a single amino acid mutation are responsible for reduced sialidase activity in mice carrying the Neu1a allele.

    abstract::The Neu1 locus, in the S region of the murine histocompatibility-2 complex, regulates the sialic acid content of several liver lysosomal enzymes. Three alleles, Neu1a, Neu1b, and Neu1c, have been described on the basis of differential sialylation of the enzyme liver acid phosphatase. The Neu1a allele occurs in a small...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.7.975

    authors: Carrillo MB,Milner CM,Ball ST,Snoek M,Campbell RD

    更新日期:1997-10-01 00:00:00

  • A new method for the analysis of amide-linked hydroxy fatty acids in lipid-As from gram-negative bacteria.

    abstract::Lipid-A represents the ubiquitous, covalently bound hydrophobic component of bacterial lipopolysaccharides (endotoxins). Lipid-As isolated and characterized from rhizobial species have large variations in their backbone sugars, as well as in their hydroxy fatty acid substituents. The sugar backbones consist of either ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/2.6.535

    authors: Bhat UR,Carlson RW

    更新日期:1992-12-01 00:00:00

  • Analysis of the specific interactions between the lectin domain of malectin and diglucosides.

    abstract::The endoplasmic reticulum malectin is a highly conserved protein in the animal kingdom that has no counterpart so far in lower organisms. We recently determined the structure of its conserved domain and found a highly selective binding to Glc(2)Man(9)GlcNAc(2), an intermediate of N-glycosylation. In our quest for puta...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwq059

    authors: Schallus T,Fehér K,Sternberg U,Rybin V,Muhle-Goll C

    更新日期:2010-08-01 00:00:00

  • Prevention of the death of the rat axotomized hypoglossal nerve and promotion of its regeneration by bovine brain gangliosides.

    abstract::We have examined the time course of the neuronal death and regeneration of rat axotomized hypoglossal nerve with various conditions of the nerve resection, and established a useful system to measure neurotrophic activities of bioactive substances. In this system, neuronal death can be evaluated by counting surviving n...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.11.1247

    authors: Itoh M,Fukumoto S,Baba N,Kuga Y,Mizuno A,Furukawa K

    更新日期:1999-11-01 00:00:00

  • C-Mannosylation of MUC5AC and MUC5B Cys subdomains.

    abstract::We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh041

    authors: Perez-Vilar J,Randell SH,Boucher RC

    更新日期:2004-04-01 00:00:00

  • Incorporation of orally applied (13)C-galactose into milk lactose and oligosaccharides.

    abstract::Biosynthesis and functions of human milk oligosaccharides (HMO) are not well known. A typical housekeeping enzyme, beta1,4-galactosyltransferase, links galactose to glucose to form lactose which is then used as backbone for the assembly of HMO. We investigated whether milk lactose and HMO may be labeled in vivo by an ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwj092

    authors: Rudloff S,Obermeier S,Borsch C,Pohlentz G,Hartmann R,Brösicke H,Lentze MJ,Kunz C

    更新日期:2006-06-01 00:00:00

  • Structural analysis of oligosaccharide-alditols released by reductive beta-elimination from oviducal mucins of Bufo bufo: characterization of the carbohydrate sequence Gal(alpha1-3)GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal.

    abstract::Several tri- to hexasaccharide-alditols of the jelly coat surrounding the eggs of Bufo bufo were studied by methylation analysis, MALDI-TOF mass spectrometry, and 1H-NMR spectroscopy. As observed for six other amphibian species, these carbohydrate chains are highly species-specific. The main characteristics of the spe...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.6.777

    authors: Morelle W,Strecker G

    更新日期:1997-09-01 00:00:00

  • A sequencing strategy for the localization of O-glycosylation sites of MUC1 tandem repeats by PSD-MALDI mass spectrometry.

    abstract::It is demonstrated with glycopeptides of the polymorphic epithelial mucin (MUC1) that post-source decay matrix-assisted laser desorption ionization (PSD-MALDI) is a fast, highly sensitive, and reproducible method for the localization of O-glycosylation sites by reflectron time-of-flight (TOF) mass spectrometry. We hav...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.7.881

    authors: Goletz S,Thiede B,Hanisch FG,Schultz M,Peter-Katalinic J,Müller S,Seitz O,Karsten U

    更新日期:1997-10-01 00:00:00

  • Molecular characterization of beta-trace protein in human serum and urine: a potential diagnostic marker for renal diseases.

    abstract::We have isolated beta-trace protein from cerebrospinal fluid, serum, plasma, and urine samples of normal volunteers and sera and hemofiltrate of patients with chronic renal failure. Blood-derived and urinary beta-trace have significantly higher molecular weights than their cerebrospinal fluid counterpart, the amino ac...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.4.499

    authors: Hoffmann A,Nimtz M,Conradt HS

    更新日期:1997-06-01 00:00:00

  • De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering.

    abstract::Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mech...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwu121

    authors: André-Miral C,Koné FM,Solleux C,Grandjean C,Dion M,Tran V,Tellier C

    更新日期:2015-04-01 00:00:00

  • Rates and impact of human antibody glycation in vivo.

    abstract::Glycation of immunoglobulin G (IgG) can result from incubation with a reducing sugar in vitro or during circulation in vivo. Upon injection of a recombinantly produced human therapeutic IgG into humans, changes in the glycation levels could be observed as a function of circulation time. Mass changes on the individual ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwr141

    authors: Goetze AM,Liu YD,Arroll T,Chu L,Flynn GC

    更新日期:2012-02-01 00:00:00

  • Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula polymorpha.

    abstract::Presently almost no information is available on the oligosaccharide structure of the glycoproteins secreted from the methylotrophic yeast Hansenula polymorpha, a promising host for the production of recombinant proteins. In this study, we analyze the size distribution and structure of N-linked oligosaccharides attache...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh030

    authors: Kim MW,Rhee SK,Kim JY,Shimma Y,Chiba Y,Jigami Y,Kang HA

    更新日期:2004-03-01 00:00:00

  • Functional expression of the capsule polymerase of Neisseria meningitidis serogroup X: a new perspective for vaccine development.

    abstract::Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis and sepsis. A key feature in pathogenicity is the capsular polysaccharide (CPS) that prevents complement activation and thus supports bacterial survival in the host. Twelve serogroups characterized by immunologically and structurally different CPSs...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwt102

    authors: Fiebig T,Berti F,Freiberger F,Pinto V,Claus H,Romano MR,Proietti D,Brogioni B,Stummeyer K,Berger M,Vogel U,Costantino P,Gerardy-Schahn R

    更新日期:2014-02-01 00:00:00

  • Comparison of the substrate specificities and catalytic properties of the sister N-acetylglucosaminyltransferases, GnT-V and GnT-Vb (IX).

    abstract::N-Acetylglucosaminlyltransferase-V (GnT-V) synthesizes GlcNAcbeta1,6Man branched N-glycans both in vitro and in vivo. A paralog, GnT-Vb (or GnT-IX), has also been shown to synthesize both GlcNAcbeta1,6Man branched N- and O-glycans. GnT-V is expressed in most human and rodent tissues while GnT-Vb expression is limited ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp158

    authors: Alvarez-Manilla G,Troupe K,Fleming M,Martinez-Uribe E,Pierce M

    更新日期:2010-02-01 00:00:00

  • Mice lacking sialyltransferase ST3Gal-II develop late-onset obesity and insulin resistance.

    abstract::Sialyltransferases are a family of 20 gene products in mice and humans that transfer sialic acid from its activated precursor, CMP-sialic acid, to the terminus of glycoprotein and glycolipid acceptors. ST3Gal-II (coded by the St3gal2 gene) transfers sialic acid preferentially to the three positions of galactose on the...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cww098

    authors: Lopez PH,Aja S,Aoki K,Seldin MM,Lei X,Ronnett GV,Wong GW,Schnaar RL

    更新日期:2017-01-01 00:00:00

  • Identification and synthesis of a novel 15 kDa beta-galactoside-binding lectin in human leukocytes.

    abstract::Knowledge of the identity, synthesis and secretion of beta-galactoside-binding lectins by leukocytes is of importance because lactosaminoglycans present at the leukocyte cell surface may be physiologically significant lectin receptors that could mediate autocrine or paracrine functions and/or cell adhesion. This paper...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/2.4.285

    authors: Sharma A,DiCioccio RA,Allen HJ

    更新日期:1992-08-01 00:00:00

  • Structural characterization of the bovine tracheal chondroitin sulfate chains and binding of Plasmodium falciparum-infected erythrocytes.

    abstract::Sequestration of Plasmodium falciparum-infected red blood cells (IRBCs) in the human placenta is mediated by chondroitin 4-sulfate (C4S). A cytoadherence assay using chondroitin sulfate proteoglycans (CSPGs) is widely used for studying C4S-IRBC interactions. Bovine tracheal chondroitin sulfate A (CSA) preparation lack...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh077

    authors: Muthusamy A,Achur RN,Valiyaveettil M,Madhunapantula SV,Kakizaki I,Bhavanandan VP,Gowda CD

    更新日期:2004-07-01 00:00:00

  • Subunit-specific sulphation of oligosaccharides relating to charge-heterogeneity in porcine lutrophin isoforms.

    abstract::Lutrophin (LH) consists of an array of isoforms with different charges and bioactivities. This study was undertaken to clarify specifically how oligosaccharides of alpha and beta subunits contribute to LH isoform charges. Porcine LH (pLH) was separated into four isoforms by isoelectric focusing (IEF), followed by subu...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/2.3.225

    authors: Ujihara M,Yamamoto K,Nomura K,Toyoshima S,Demura H,Nakamura Y,Ohmura K,Osawa T

    更新日期:1992-06-01 00:00:00