Lectins, carbohydrate-immobilized

Recently, Vomholt et al. reported a cuvette-based SPR method for examining binding domains of lectins on carbohydrate-immobilized surfaces [19]. By taking advantage of the low sample volume required by their cuvette-based method. 

Crude chloroform-methanol-water (30 60 8, v/v) extracts of immunostainedTLC bands were analyzed without further purification by nanoelectrospray low-energy mass spectrometry. The authors showed that this effective PLC/MS-joined procedure offers a wide range of applications for any carbohydrate-binding agents such as bacterial toxins, plant lectins, and others. Phenyl-boronic acid (PBA) immobilized on stationary support phases can be put to similar applications. This technology, named boronate affinity chromatography (BAC), consists of a chemical reaction of 1,2- and 1,3-diols with the bonded-phase PBA to form a stable... 

Schmidt and co-workers have also immobilized the mannose derivatives to the commercially available glass slides under reductive amination or acylation conditions [167], Their study showed that the ConA interaction was in accordance with those obtained in aqueous solution and other immobilization methods, and non-specific protein binding to the functionalized glass slides was negligible. The same carbohydrate microarray plate could also be tested for other lectins, therefore the precious immobilized carbohydrates can be reused. 

CRP binds terminal -galactosides at a pH optimum of 5-6 and lectin binding assays require that both lectin and ligand be immobilized. Thus it has been suggested that semm CRP does not function as a lectin under normal physiological conditions and binds carbohydrate only under conditions where tissues are inflamed or hypoxic [208]. 

Many of the techniques used to enrich PTMs at the protein level are applicable at the peptide level. A popular method for enrichment of phosphopeptides is to use an immobilized metal affinity column. The molecular basis for the enrichment is the phosphate affinity to transition metal ions, such as copper, nickel, cobalt, iron, aluminum, gallium, and zinc (142). A comprehensive source for phosphopeptide enrichment strategies can be found in a recent review (143). Lectin-based chromatography is used to enrich for glycopep-tides (144). Lectins are proteins that have an affinity for carbohydrates. With multiple available enrichment options, choosing the right one for the experiment is important. For determining which proteins have a certain PTM, protein-level enrichment can be performed. However, if a type of PTM is of... 

Firstly, immobilized lections may be used. Lectins are proteins that display high binding selectivity for defined carbohydrate structures and sugar residues [23, 24]. The interaction with the glycosylated enzyme is non-covalent, and binding strength depends on the specific association constants. This approach is cost intensive because it affords the production of a further immobilized protein. 

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