Abstract
Neuronal growth inhibitory factor (GIF) is first found in theAlzheimer's disease related study which shows a unique functionin inhibiting the survival and outgrowth of cortical neuron.Primary structure of GIF reveals a high homology withmetallothionein (MT) family. GIF gene was further demonstratedto be located at the same chromosome as that of MT gene. GIF istherefore designated as MTIII.To explore the differentialfunction between MTII and MTIII, structural analysis isrequired. For this purpose, purification of MTIII in largequantity is necessary. Attempts have been made to purify theprotein by conventional column chromatographies. However, therecovery and purity was not as good as expected. We tried to usereverse phase HPLC column to improve the purification andobtained a high yield and purified MTIII. The size of theprotein was confirmed by mass spectrometry. The purified proteinwas used to analyze the spectroscopic properties which includeultraviolate absorption and circular dichroism. The resultsdemonstrated that both MT and MTIII have similar metal-thiolatecharacteristics. In addition, protein was analyzed by cadmiumand proton 1D-NMR. Partial sequential resonance assignment andsecondary structure in Cd-GIF have obtained.