Synthesis of the C-terminal domain of the tissue inhibitor of metalloproteinases-1 (TIMP-1). |
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Authors: | József Bódi Nikolett Mihala Andrea Hajnal Katalin F Medzihradszky Helga Süli-Vargha |
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Institution: | Research Group for Peptide Chemistry, Hungarian Academy of Sciences, E?tv?s Loránd University, H-1518 Budapest 112 PO Box 112, Hungary. |
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Abstract: | According to recent investigations, the C-terminal domain of the tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) is responsible for some biological effects that are independent of the enzyme-inhibiting effect of the N-terminal domain of the molecule. The C-terminal domain has been prepared for structure-biological activity investigations. After the chemical synthesis and the folding of the linear peptide. LC-MS and MALDI-MS analysis revealed that two isomers with different disulphide bond arrangements were formed. Since more than 30 folding experiments resulted in products with a very similar HPLC-profile, it was concluded that in the absence of the TIMP-1 N-terminal domain no entirely correct folding of the C-terminal domain occurred. Furthermore, it was observed that, in spite of several purification steps, mercury(II) ions were bound to the 6SH-linear peptide; it was demonstrated--using disulphide bonded TIMP-1(Cys145-Cys166) as a model--that mercury(II) ions can cause peptide degradation at pH 7.8 as well as in 0.1% trifluoroacetic acid. |
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Keywords: | TIMP‐1 C‐terminal domain synthesis structural isomers mass spectrometry disulphide‐bridge mercury(II) ions degradation |
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