Essentials of Carbohydrate Chemistry

Robyt, John F.

doi:10.1007/978-1-4612-1622-3

Cited by 239 publications

(236 citation statements)

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“…Pullulan is a highly soluble, linear polymer comprised of repeating maltotriose and maltotetraose units (27). Protein concentrations in the micromolar range were required to observe significant binding to the pullulan-coupled sensor chip; examples of the sensorgrams obtained using the pullulan-coupled chip are given in Fig.…”

Section: Fig 5 Comparison Of Off-rates By Sprmentioning

confidence: 99%

Insights into the Conformational Equilibria of Maltose-binding Protein by Analysis of High Affinity Mutants

Telmer

Shilton

2003

Journal of Biological Chemistry

106

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ATP binding cassette transporters couple ATP hydrolysis to the transmembrane transport of a diverse range of compounds. Members of the ATP binding cassette transporter superfamily are characterized by two membrane-integral domains that each contain 6 or more membrane spanning helices, but are otherwise poorly conserved, and two peripheral ATP binding cassette domains that display sequence conservation across the entire superfamily (1). In addition to the membrane complex, ATP binding cassette systems that catalyze nutrient uptake have primary receptors (binding proteins) that serve two functions: they provide a high affinity binding site for the transported molecule and they regulate the ATPase activity of the integral membrane complex.We are interested in the function of the primary receptors in the transport process. As a group, these proteins have been intensively studied by x-ray crystallography and other biophysical techniques (for a review, see Ref.2). They typically contain two domains separated by a hinge region; the substrate binds in the cleft between the two domains, and the protein undergoes a large conformational change, leading to closure of the cleft. With respect to the maltose transport system, domain closure in the binding protein is thought to be the first step toward molecular shape recognition by the membrane complex (3, 4), although it has been shown that both substrate-loaded and substrate-free binding proteins have a role in the transport cycle (5-8). The association and dissociation of the substrate, and attendant conformational changes in the binding protein (MBP), 1 may have direct effects on transport kinetics and regulation of ATP hydrolysis by MalFGK 2 . To investigate the role of binding protein affinity on the transport process, our goal was to engineer MBP molecules with greater affinity for maltose, without changing residues in either the maltose binding site or in regions thought to interact with MalFGK 2 .Crystal structures of MBP in both the closed and open conformations have been solved (9, 10), and they show that binding of maltose results in a large conformational change of the protein, bringing the two domains together such that the substrate is buried inside the cleft. In solution, unliganded MBP is in the open conformation (11); however, there is no obvious energetic barrier to closure of the ligand binding cleft, either in the hinge or in the interface surrounding the ligand binding site. Rather, an interface on the opposite side of the hinge from the ligand binding site appears to maintain the protein in an * This work was supported by Natural Sciences and Engineering Research Council Grant 21749-1999 (to B. H. S.); the macromolecular x-ray facility at the University of Western Ontario was financed with grants from the Canada Foundation for Innovation, Ontario Challenge Fund, and Western's Academic Development Fund; fluorescence and SPR studies were carried out at the University of Western Ontario Biomolecular Interactions and Conformations Facility, supported by a Mu...

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Section: Fig 5 Comparison Of Off-rates By Sprmentioning

confidence: 99%

Insights into the Conformational Equilibria of Maltose-binding Protein by Analysis of High Affinity Mutants

Telmer

Shilton

2003

Journal of Biological Chemistry

106

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“…Chitin is the most abundant glycopolymer constituting the integument including the shells of crustaceans, insect exoskeleton, microfilarial sheath of parasitic nematodes, cell wall of most fungi, and spore shell of microsporidia [4,7,16]. Chitinases are ubiquitous chitin-fragmenting hydrolases and produced by a vast array of organisms [5].…”

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confidence: 99%

Quantitative Changes in Serum Concentration of Bovine Gut Chitinase in Theileria Infection

Fujimoto

Kimura

Suzuki

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. Bovine gut chitinase is exclusively produced in the liver and secreted into the blood. In the present study, we established a semi-quantitative method by Western blot analysis for measurement of the chitinase content in blood and examined its alteration during postnatal development and experimental infection with hemoprotozoan parasite in cattle. Its serum levels from 1 week to 1 year of age showed a significant increase only in 3-4-month-old group. The plasma concentration of the gut chitinase was not changed during acute inflammation caused by lipopolysaccharide but increased gradually after a Theileria injection and peaked at 52 days post-infection. It appears that the increase in the blood chitinase levels might be a defensive response in cattle against protozoan infection. KEY WORDS: bovine serum, chitinase, Theileria sergenti.

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“…Since the tag was originally published [8], the indicated minimal structural motif has been discovered. D8 tag was shown to bind specifically to Sephadex G-100 (Pharmacia), which is a commonly used gel filtration matrix, produced by crosslinking of dextran B512 with epichlorohydrin to form beads [16,17]. In contrast, the D8 tag does not bind to other similar matrices such as Sepharose and Seph-acryl.…”

Section: The Motifs That Bind Sephadex and Streptavidinmentioning

confidence: 99%

“…This is an important consideration when combining a Sephadex affinity step with additional steps that might involve other types of chromatography resin or solid supports. The binding of the tag to Sephadex can be efficiently competed with dextran B512, which is a homopolymer of glucose connected mainly via α-1,6 glucosidic linkages (95%) and occasional α-1,3 linkages (5%) [16]. Although the D8 tag was originally selected using Sephadex G-100 as a target, it can bind Sephadex G-200 with equal efficiency.…”

Section: The Motifs That Bind Sephadex and Streptavidinmentioning

confidence: 99%

RNA affinity tags for purification of RNAs and ribonucleoprotein complexes

Srisawat

Engelke

2002

Methods

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Intrinsic affinity tags are useful tools for the study of macromolecular targets. Although polypeptide affinity tags are routinely used in purification and detection of protein complexes, there has been a relative lack of powerful RNA affinity tags that can be embedded within RNA sequences. Here, the preparation and use of two RNA affinity tags against Sephadex or streptavidin are described. The two tags have different strengths that make them appropriate for slightly different uses. One is a high-affinity ligand for streptavidin that can be specifically eluted by competition with biotin under otherwise native binding conditions. The other tag binds selectively to Sephadex beads, and can be eluted by competition with the soluble dextran that composes Sephadex. When properly placed within another RNA molecule, the tags can be used to effect dramatic purification of RNA or ribonucleoprotein complexes from complex mixtures of cellular RNA.

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Essentials of Carbohydrate Chemistry

Cited by 239 publications

References 0 publications

Insights into the Conformational Equilibria of Maltose-binding Protein by Analysis of High Affinity Mutants

Insights into the Conformational Equilibria of Maltose-binding Protein by Analysis of High Affinity Mutants

Quantitative Changes in Serum Concentration of Bovine Gut Chitinase in Theileria Infection

RNA affinity tags for purification of RNAs and ribonucleoprotein complexes

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