Lessons on the critical interplay between zinc binding and protein structure and dynamics

Zinc is one of the most important micronutrients for virtually all living organisms, and hence, it is important to understand the molecular mechanisms for its homeostasis. Besides proteins involved in transmembrane transport, both extra- and intracellular zinc-binding proteins play important roles in the respective metabolic networks. Important examples for extracellular zinc transporters are mammalian serum albumins, and for intracellular zinc handling, certain metallothioneins are of relevance. The availability of protein structures including relevant metal binding sites is a fundamental prerequisite to decipher the mechanisms that govern zinc binding dynamics in these proteins, but their determination can prove to be surprisingly challenging. Due to the spectroscopic silence of Zn2 +, combinations of biophysical techniques including electrospray ionisation mass spectrometry (ESI-MS) and multinuclear NMR, isothermal titration calorimetry (ITC) and extended X-ray absorption fine structure (EXAFS) spectroscopy, coupled with site-directed mutagenesis and molecular modelling have proven to be valuable approaches to understand not only the zinc-binding properties of metallothioneins and albumins, but also the influence of other physiologically relevant competing agents. These studies have demonstrated why the bacterial metallothionein SmtA contains a site inert towards exchange with Cd2 +, why the plant metallothionein EC from wheat is partially unfolded in the presence of Cd2 +, and how fatty acids impact on the zinc-binding ability of mammalian serum albumins.

Understanding the molecular mechanisms that govern zinc movements in biological systems requires the study of structures of relevant proteins, their zinc binding sites, and how metal binding and structural dynamics including conformational changes affect each other.Figure optionsDownload as PowerPoint slideHighlights
► The bacterial metallothionein SmtA contains a zinc finger fold.
► SmtA contains a site inert towards metal exchange.
► EC MT can only form an ordered fold in the presence of Zn, but not Cd.
► Zinc binding by albumin is allosterically linked to fatty acid binding.