On the basis of hplc it can be shown that the major

On the basis of HPLC it can be shown that the major
cannabinoids are specifically accumulating after a shorten-
ing of the light period. Chemotypes are additionally con-
firmed using qPCR specific for thca and cbda genes. Gene
transcript based chemotyping was only possible comparing
the thca and cbda transcription level ratio at the middle of
the cultivation period, while at the end differences in gene
transcription ratio diminished. 1H-NMR results indicate a
clear difference between both chemotypes and cultivation
phases by applying pattern recognition software.
Figure 1. Ratio analysis of the metabolites (Δ9-THCA/
CBDA) and the relative transcription between cbda and thca
(2-(ΔΔCt), Y-ax (right) in logarithm), shown per C. sativa L.
variety. A) Represent ratio analysis of chemotype I C. sativa
L. var. Bedrobinol. B) Represent ratio analysis of chemo-
type II C. sativa L. var. Bediol. No detectable transcription
ratio could be obtained for the chemotype I C. sativa L. var.
Bedrobinol in week 2 and 3 due to an absence of cbda tran -
scripts. Error bars are defined by upper and lower limits for
gene transcription and by SD for metabolites.
L2.6
RAGE receptor dimeric structure studied
by hydrogen-deuterium exchange
monitored by mass spectrometry
Ewa Sitkiewicz, Krzysztof Tarnowski, Magda
Kulma, Jarosław Poznański, Michal Dadlez
Institute of Biochemistry and Biophysics PAS, Warszawa, Poland
e-mail: Michal Dadlez <[email protected]>
Introduction : Receptor for advanced glycation end prod-
ucts (RAGE) is a multiligand cell surface receptor, known
to bind glycoxidised form of proteins as well as many other
ligands such as A‐ peptide, the S100 family of proinflam-
matory cytokine-like mediators, Mac1 integrin and many
others. RAGE is an important mediator of the proinflam-
matory response involved in diverse pathophysiological
states such as neurological disorders, stroke, amyloidosis,
immune response, diabetes and inflammatory disorders,
infectious disease and tumors. It is a potential therapeutic
target, but in spite of significant effort its structure, ligand
binding mode and signal transduction pathway has not
been characterised yet on the molecular level.
Methods : In the presented work we used hydrogen-deute-
rium exchange and mass spectrometry to gain insight into
the structural properties of exRAGE — the full extracel-
lular part of the protein containing V, C1 and C2 domains
and the structural consequences of its dimerisation.
Results : We have identified two regions in RAGE se-
quence which undergo protection when the protein dimer-
ises. These regions map to the region between domains C1
and C2, indicating the molecular mechanism of stabilisa-
tion of a flexible linker region. Based on this we propose a
novel model of RAGE dimer and tetramer.