Bionanalytical Core Unit

The structural and functional knowledge of natural hydrogels of the lung and intestine is sparse. This is largely caused by the enormous complexity of the underlying molecular networks that are composed of large protein chains, which are heavily glycosylated at multiple sites. Although it is accepted that the barrier function of the mucus is of fundamental importance to maintain the healthy state, a thorough and detailed analysis is required to unravel the structure-function relationship. Within this CRC, this complexity will be addressed using multiple orthogonal analytical techniques.

Lung and intestine derived mucin samples of healthy and diseased states (mouse and human) and from cell culture models will be characterized using a multi-step approach consisting of: 1) the production and harvesting of glycoproteins, 2) proteomics approaches to characterize and sequence mucin proteins, 3) glycomics approaches to identify characteristic glycosylation features and 4) glycoproteomics techniques to elucidate the distribution of relevant glycan structures along the protein backbone. In addition, mass-spectrometry based approaches will be complemented by 5) state of the art electron microscopy and electron tomography techniques to analyze mucus glycoproteins and their assemblies in their native, solvated state. Some of the utilized techniques are readily available, while others will be developed and subsequently transferred to this project.