Sylabus
Lectures
The historical development of understanding of hormonal receptors (since Paul Ehrlich and John Newport Langley)
Discovery of adenylylcyclase and cAMP (Earl Sutherland) = the key stone of molecular endocrinology and pharmacology
Membrane receptors ? classification; ligand, agonist, antagonist, partial agonist, inverse agonist
Radioisotopes in medicine and biology, liquid scintillation, beta and gama counting, Bq, Ci, radioligand, specific radioactivity, chemical stability of the radioligand, radiochemical stability
Ligand binding studies of membrane receptors, specific radioligands, saturation binding isotherm, Scatchard plot, calculation of Bmax and Kd values, allosteric and co-operative models of receptor-ligand interactions, Hill coefficient; criteria of Pedro Cuatrecassas for identification of receptor with the specific radioligand binding site
Why there are so many receptors for the single hormone or neurotransmitter? Heterogeneity of G protein coupled receptors (GPCR), splicing variants of 7TM receptors
Trimeric GTP binding regulatory proteins (G-proteins), discovery of trimeric G proteins (Martin Rodbell and Alfred G. Gilman)
Classification of the five main families of trimeric G proteins
Phosphoplipases C, phospholipases A2, phosphodiesterases and secondary messengers
Desensitisation of hormone response, internalisation, recycling and down-regulation of GPCR; internalisation and down-regulation of trimeric G proteins, cross-talk phenomena, membrane domains (Neubig, R. R. (1994) Membrane organisation in G-protein mechanism. FASEB J. 8, 939-946), feed-back regulations, multiple signalling cascades initiated by the same hormone or neutrotransmitter.
Sodium plus potassium activated, magnesium dependent adenosine triphosphatase (Na,K-ATPase), sodium pump, [3H]ouabain as specific marker of plasma membranes
Subcellular fractionation by differential and density-gradient centrifugation; isolation of plasma membranes and other subcellular particles; plasma membrane markers and enzyme activities.
Anotace
Cílem přednášky je poskytnout přehled o základech molekulární farmakologie, základního vědeckého výzkumu, který charakterizuje hormonální receptory a jejich signální kaskády s pomocí metod biochemie, farmakologie a buněčné fyziologie. Důraz je kladen na postižení postupného vývoje poznání.