Height determination techniques for the next national height system of Finland - a case study

Precise levelling is known for its accuracy and reliability in height determination, but the process itself is slow, laborious and expensive. We have started a project to study methods for height determination that could decrease the creation time of national height systems without losing the accuracy and reliability that is needed for them. In the pilot project described here, we study some of the alternative techniques with a pilot field test where we compared them with the precise levelling. The purpose of the test is not to evaluate the mutual superiority or suitability of the techniques, but to establish the background for a larger test and to find strong and weak points of each technique. The techniques chosen for this study were precise levelling, Mobile Laser Scanning (MLS) and Global Navigation Satellite System (GNSS) levelling, which included static Global Positioning System (GPS) and Virtual Reference Station (VRS) measurements. This research highlighted the differences of the studied techniques and gave insights about the framework and procedure for the later experiments. The research will continue in a larger scale, where the suitability of the techniques regarding the height systems is to be determined

Alternative conformations of the x region of human protein disulphide-isomerase modulate exposure of the substrate-binding b{'} domain

Protein disulphide isomerase (PDI) is a key multi-domain protein folding catalyst in the endoplasmic reticulum. The b' domain of PDI is essential for the non-covalent binding of incompletely folded protein substrates. Earlier, we defined the substrate binding site in the b' domain of human PDI by modelling and mutagenesis studies. Here, we show by fluorescence and NMR that recombinant human PDI b'x (comprising the b' domain and the subsequent x linker region) can assume at least two different conformations in solution. We have screened mutants in the b'x region to identify mutations that favour one of these conformers in recombinant b'x, and isolated and characterised examples of both types. We have crystallised one mutant of b'x (I272A mutation) in which one conformer is stabilized, and determined its crystal structure to a resolution of 2.2 A. This structure shows that the b' domain has the typical thioredoxin fold and that the x region can interact with the b' domain by "capping" a hydrophobic site on the b' domain. This site is most likely the substrate binding site and hence such capping will inhibit substrate binding. All of the mutations we previously reported to inhibit substrate binding shift the equilibrium towards the capped conformer. Hence, these mutations act by altering the natural equilibrium and decreasing the accessibility of the substrate binding site. Furthermore, we have confirmed that the corresponding structural transition occurs in the wild type full-length PDI. A cross-comparison of our data with that for other PDI-family members, Pdi1p and ERp44, suggests that the x region of PDI can adopt alternative conformations during the functional cycle of PDI action and that these are linked to the ability of PDI to interact with folding substrates