Electrophoretic Behavior of Copolymeric Galacturonans Including Comments on the Information Content of the Intermolecular Charge Distribution

Williams, Martin A. K.; Cucheval, A.; Ström, Anna; Ralet, Marie-Christine

doi:10.1021/bm900119u

Cited by 21 publications

(25 citation statements)

References 73 publications

(85 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…27,28 These pectins were named as: 37R, 37B, 58R, 58B, 75R and 86R, where the numbers indicate the degree of methyl-esterification, and the letters 'R' and 'B' indicate random and block distribution pattern of the uncharged methyl-ester groups. Pectins with 75% degree of methyl-esterification and 86% degree of methylesterification were purchased from Sigma Aldrich (Missouri, US).…”

Section: Methodsmentioning

confidence: 99%

Structural mechanism of complex assemblies: characterisation of beta-lactoglobulin and pectin interactions

Melton

Jameson

et al. 2015

Soft Matter

Self Cite

View full text Add to dashboard Cite

Knowledge of how proteins and polysaccharides interact is the key to understanding encapsulation and emulsification in these composite systems and ultimately to understanding the structures of many biological network systems. As a model system we have studied β-lactoglobulin A (βLgA) interacting with pectins of various amounts and distribution patterns of charge. The studies were conducted at pH 4 at minimal ionic strength, where the βLgA and the pectins are oppositely charged, resulting in an electrostatic attraction to each other. Isothermal titration calorimetry (ITC) experiments were performed to determine the thermodynamics associated with βLgA-pectin titration. It was found that βLgA only interacted with pectins with an adequate amount of charge, and that the complexation between βLgA and pectin was a two-step process initially involving binding of the protein to available sites on the pectin, and subsequently binding of the protein onto the bound protein that has previously adsorbed. Circular dichroism (CD) and intrinsic tryptophan fluorescence were also measured of βLgA during its interaction with the pectin samples, and show that the binding leads to significant conformational changes in βLgA. An increase in the turbidity of the solution of the resultant complexes indicates the formation of large-scale interpolymer associations of the primary complexes mediated by protein-rich domains.

show abstract

Section: Methodsmentioning

confidence: 99%

Structural mechanism of complex assemblies: characterisation of beta-lactoglobulin and pectin interactions

Melton

Jameson

et al. 2015

Soft Matter

Self Cite

View full text Add to dashboard Cite

show abstract

“…Pectin (Mw 30-100 kDa), extracted from apple pomace, was purchased from Fluka Biochemika (Sigma Aldrich, Switzerland) as a starting material. The sample average DM and the degree of blockiness [32] of this sample, and that produced by modification as described below, were determined using capillary zone electrophoresis as previously described [39,40].…”

Section: Pectinmentioning

confidence: 99%

Micro-rheological behaviour and nonlinear rheology of networks assembled from polysaccharides from the plant cell wall

Vincent¹,

Mansel

Kramer

et al. 2013

New J. Phys.

View full text Add to dashboard Cite

The same fundamental questions that have driven enquiry into cytoskeletal mechanics can be asked of the considerably less-studied, yet arguably just as important, biopolymer matrix in the plant cell wall. In this case, it is well-known that polysaccharides, rather than filamentous and tubular protein assemblies, play a major role in satisfying the mechanical requirements of a successful cell wall, but developing a clear structure-function understanding has been exacerbated by the familiar issue of biological complexity. Herein, in the spirit of the mesoscopic approaches that have proved so illuminating in the study of cytoskeletal networks, the linear microrheological and strain-stiffening responses of biopolymeric networks reconstituted from pectin, a crucial cell wall polysaccharide, are examined. These are found to be well-captured by the glassy worm-like chain (GWLC) model of self-assembled semi-flexible filaments. Strikingly, the nonlinear mechanical response of these pectin networks is found 6

show abstract

“…HG-93 aliquots were deesteriWed by adequate amounts of 0.2 M NaOH to yield HG-B69 (base-deesteriWed HG, DM 69) and HG-B20 (base-deesteriWed HG, DM 20). Base-deesteriWcation led to a random repartition of charges (Williams et al 2009). A HG-93 aliquot was deesteriWed using plant-PME to yield HG-P36 (plant-PME-deesteriWed HG, DM 36), plant-PME deesteriWcation leading to a blockwise repartition of charges (Ralet et al 2001).…”

Section: Polysaccharide Isolationmentioning

confidence: 99%

Monoclonal antibodies to rhamnogalacturonan I backbone

Ralet

Tranquet

Poulain

et al. 2010

Planta

115

View full text Add to dashboard Cite

Monoclonal antibodies were raised against rhamnogalacturonan I backbone, a pectin domain, using Arabidopsis thaliana seed mucilage-derived rhamnogalacturonan I oligosaccharides--BSA conjugates. Two monoclonal antibodies, designated INRA-RU1 and INRA-RU2, selected for further characterization, were specific for the backbone of rhamnogalacturonan I, displaying no binding activity against the other pectin domains i.e. homogalacturonans, galactans or arabinans. A range of oligosaccharides was prepared by enzymatic digestion of rhamnogalacturonan I isolated from Arabidopsis thaliana seed mucilage and from sugar beet pectin, purified by low-pressure chromatography and characterized by high-performance anion-exchange chromatography and mass spectrometry. These rhamnogalacturonan I oligomers were used to characterize the binding site of the two monoclonal antibodies by competitive inhibition. Both INRA-RU1 and INRA-RU2 showed maximal binding to the [-->2)-alpha-L-rhamnosep-(1-->4)-alpha-D-galacturonic acid p-(1-->](7) structural motif but differed in their minimum binding requirement. INRA-RU2 required at least two disaccharide (rhamnose-galacturonic acid) repeats for the antibody to bind, while INRA-RU1 required a minimum of six disaccharide repeats. Furthermore, the binding capacity of INRA-RU1 decreased steeply as the number of disaccharide repeats go beyond seven. Each of these antibodies reacted with hairy regions isolated from sugar beet pectin. Immunofluorescence microscopy indicated that both antibodies can be readily used to detect rhamnogalacturonan I epitopes in various cell wall samples.

show abstract

Electrophoretic Behavior of Copolymeric Galacturonans Including Comments on the Information Content of the Intermolecular Charge Distribution

Cited by 21 publications

References 73 publications

Structural mechanism of complex assemblies: characterisation of beta-lactoglobulin and pectin interactions

Structural mechanism of complex assemblies: characterisation of beta-lactoglobulin and pectin interactions

Micro-rheological behaviour and nonlinear rheology of networks assembled from polysaccharides from the plant cell wall

Monoclonal antibodies to rhamnogalacturonan I backbone

Contact Info

Product

Resources

About