The carbohydrate recognition domain of collectins

Collectins are effector molecules of the innate immune system that play an important role in the first line of defence against bacteria, viruses and fungi. Most of their interactions with microorganisms are mediated through their carbohydrate recognition domain (CRD), which binds in a Ca(2+)-dependent manner to glycoconjugates. This domain is a well-known structure that is present in a larger group of proteins comprising the C-type lectin domain family. Collectins form a subgroup within this family based on the presence of a collagen domain and the trimerization of CRDs, which are essential for the ligand-binding properties of these proteins. The ligand specificity among the nine collectin members is significantly different as a result of both the structural organization of the trimers and specific sequence changes in the binding pocket of the CRD. In addition, some collectin members have additional features, such as N-linked glycosylation of CRD residues and additional loop structures within the CRD that have a large impact on their interaction with the glycoconjugates present on microorganisms or host cells. The availability of crystal structures of three members of the collectin family (surfactant proteins A and D and mannan-binding protein) provides an important tool for addressing the impact of these CRD differences on ligand binding. In this review, the structural differences and similarities between the CRDs of collectins are summarized and their relationship with their ligand-binding characteristics is discussed.     

The complex network of non-cellulosic carbohydrate metabolism

Partitioning of carbon dominates intracellular fluxes in both photosynthetic and heterotrophic plant tissues, and has vast influence on both plant growth and development. Recently, much progress has been made in elucidating the structures of the biosynthetic and degradative pathways that link the major and minor pools of soluble carbohydrates to cellular polymers such as starch, heteroglycans and fructans. In most cases, the regulatory properties of these pathways have been elucidated and the enzymes involved have been investigated using reverse genetics approaches. Although many of the results from these approaches were merely confirmatory, several of them were highly unexpected. The challenge ahead is to achieve better understanding of metabolic regulation at the network level in order to develop more rational strategies for metabolic engineering.     

The utility of carbohydrate microarrays in glycomics

Carbohydrate microarrays are powerful tools in glycomics. Interactions of different carbohydrate structures with a wide variety of biological targets, including proteins, RNA, viruses, and whole cells, have been investigated using this technique. Binding preferences and specificities, inhibition of interactions, enzymatic activities, and structure-function relationships have been determined. Screening and characterization of antibodies have been conducted using microarrays. Binding of whole cells to the arrays has been exploited to search for novel binding proteins and to detect bacteria in blood. Here, we review the different techniques for carbohydrate microarray production and application. To illustrate the utility of arrays for glycomics research, some select experiments are discussed in greater detail.