By Sonal Srikanth, Yousang Gwack (auth.), Claus W. Heizmann (eds.)
Amajor course in scientific learn resulting in medical functions pursuits the legislation of intracellular calcium and many of the human ailments linked to an altered homeostasis of this worldwide moment messenger. those ailments contain, for instance: cardiomyopathy, irritation, mind problems, diabetes and melanoma. In Calcium-Binding Proteins and RAGE:from Structural fundamentals to ClinicalApplications,expert researchers within the box aspect a number of the equipment that are now frequent to check calcium binding proteins. those equipment and methods, resembling calcium-measurements, screening equipment, medical chemistry, and remedy, are typically appropriate to many different parts of easy and clinical examine in addition to to diagnostics. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, simply reproducible laboratory protocols, and key tips about troubleshooting and heading off identified pitfalls.
Authoritative and practical,Calcium-Binding Proteins and RAGE:from Structural fundamentals to ClinicalApplications underlines the diagnostic and medical value of this relatives of proteins in human ailments and as drug targets.
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Additional resources for Calcium-Binding Proteins and RAGE: From Structural Basics to Clinical Applications
Example text
Cox Trichloroacetic acid is from Calbiochem. The contaminating Ca2+, Mg2+, or Zn2+ was measured by atomic absorption spectrophotometry (Perkin Elmer 2380). 2. Equilibrium Dialysis Equlibrium dialysis of 1 ml protein solution was performed for 48 h in the cold room in Spectra/Por dialysis tubes (Spectrum Europe B. ) versus 200 ml buffer in polypropylene bottles on a lab shaker with one buffer change after 24 h of dialysis. 3. Fluorescent Ion Indicators Benzothiazole coumarin (BTC) and Mag-fura-2 are from Molecular Probes, Eugen, Oregon.
S100 Proteins S100 proteins (soluble in 100% ammonium sulfate) are dimeric proteins of ca 20 kDa. Each monomer contains 2 EF-hands: the C-terminal EF-hand displays the canonical sequence, whereas the N-terminal EF-hand, consisting of 14 amino acids, is specific for S100 proteins. To date, some 21 different proteins, which display various degrees of amino acid sequence homology, have been assigned to the S100 protein family (19). The protein contains a dimerization domain, an X-type four-helix bundle consisting of the N-terminal helix of EF-I and the C-terminal helix of EF-II.
4 ml. Every 2-min increments of Ca2+ are added so that the total Ca2+ concentrations in the upper compartment vary according to an exponential scale until a value of 2–3 mM is attained. The samples 2 Divalent Cation-Protein Interactions 29 of the fraction collector are supplemented with scintillation liquid and counted to 98% precision. This method has allowed us to determine quite easily and precisely the Ca2+ affinity of a great number of proteins with KCa values varying from 5 × 107 to 2 × 103 M−1.