Isolation of GIF from porcine brain and studies of its zinc transfer kinetics with apo-carbonic anhydrase |
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Authors: | Yan-bo Shi Liang Du Wei-juan Zheng Wen-xia Tang |
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Institution: | (1) State Key Laboratory of Coordination Chemistry, Coordination Chemistry Institute, Nanjing University, Nanjing, 210093, China;(2) State Key Laboratory of Pharacentical Biothnology, Department of Biochemistry, Nanjing University, Nanjing, 210093, China |
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Abstract: | Neuronal growth inhibitory factor (GIF) of porcine brain, was isolated and purified by a similar procedure which was used on the isolation of human and bovine GIF. The native porcine protein with stoichiometry of 4Cu+, 3Zn2+ was obtained for the first time. The kinetics of zinc transfer from Cu4Zn3MT-3 to apo-carbonic anhydrase were studied, and zinc transfer rate constants and thermodynamic parameters were obtained. It is found that like other MTs, porcine Cu4Zn3MT-3 can also transfer its zinc atom to apoCA, even much easier than other MTs. A possible association mechanism has been proposed, the formation of Cu4Zn3MT3-apoCA complex may be the rate-determining step. The obtained data indicate besides its neuronal growth inhibitory function, GIF might play a role in cellular Zn homeostasis in brain. |
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Keywords: | carbonic anhydrase (CA) GIF (Cu4Zn3MT-3) zinc transfer kinetic |
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