Structure and Dynamics of the N-Terminal Domain of the Cu(I) Binding Protein CusB

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• 作者：Melek N. Ucisik ; Dhruva K. Chakravorty ; Kenneth M. Merz ; Jr.
• 刊名：Biochemistry
• 出版年：2013
• 出版时间：October 1, 2013
• 年：2013
• 卷：52
• 期：39
• 页码：6911-6923
• 全文大小：696K
• 年卷期：v.52,no.39(October 1, 2013)
• ISSN：1520-4995

文摘

CusCFBA is one of the metal efflux systems in Escherichia coli that is highly specific for its substrates, Cu(I) and Ag(I). It serves to protect the bacteria in environments that have lethal concentrations of these metals. The membrane fusion protein CusB is the periplasmic piece of CusCFBA, which has not been fully characterized by crystallography because of its extremely disordered N-terminal region. This region has both structural and functional importance because it has been experimentally proven to transfer the metal by itself from the metallochaperone CusF and to induce a structural change in the rest of CusB to increase Cu(I)/Ag(I) resistance. Understanding metal uptake from the periplasm is critical to gain insight into the mechanism of the whole CusCFBA pump, which makes resolving a structure for the N-terminal region necessary because it contains the metal binding site. We ran extensive molecular dynamics simulations to reveal the structural and dynamic properties of both the apo and Cu(I)-bound versions of the CusB N-terminal region. In contrast to its functional companion CusF, Cu(I) binding to the N-terminus of CusB causes only a slight, local stabilization around the metal site. The trajectories were analyzed in detail, revealing extensive structural disorder in both the apo and holo forms of the protein. CusB was further analyzed by breaking the protein up into three subdomains according to the extent of the observed disorder: the N- and C-terminal tails, the central beta strand motif, and the M21鈥揗36 loop connecting the two metal-coordinating methionine residues. Most of the observed disorder was traced back to the tail regions, leading us to hypothesize that the latter two subdomains (residues 13鈥?5) may form a functionally competent metal-binding domain because the tail regions appear to play no role in metal binding.