Helmholtz Centre for Polar and Marine Research, Alfred Wegener Institute, Bremerhaven, Germany.
Hanyang University, Seoul, Republic of Korea.
Methods Mol Biol. 2020;2156:289-302. doi: 10.1007/978-1-0716-0660-5_20.
Several species of polar microalgae are able to live and thrive in the extreme environment found within sea ice, where ice crystals may reduce the organisms' living space and cause mechanical damage to the cells. Among the strategies adopted by these organisms to cope with the harsh conditions in their environment, ice-binding proteins (IBPs) seem to play a key role and possibly contribute to the success of microalgae in sea ice. Indeed, IBPs from microalgae predominantly belong to the so-called "DUF 3494-IBP" family, which today represents the most widespread IBP family. Since IBPs have the ability to control ice crystal growth, their mechanism of function is of interest for many potential applications. Here, we describe methods for a classical determination of the IBP activity (thermal hysteresis, recrystallization inhibition) and further methods for protein activity characterization (ice pitting assay, determination of the nucleating temperature).
几种极地微藻能够在海冰中存在并茁壮成长，在海冰中，冰晶可能会减少生物的生存空间，并对细胞造成机械损伤。在这些生物为适应环境的恶劣条件而采取的策略中，冰结合蛋白（IBP）似乎起着关键作用，并可能有助于微藻在海冰中的成功。事实上，微藻的 IBP 主要属于所谓的“DUF3494-IBP”家族，该家族目前是最广泛的 IBP 家族。由于 IBP 具有控制冰晶生长的能力，因此它们的功能机制对许多潜在的应用具有重要意义。在这里，我们描述了经典的 IBP 活性测定方法（热滞、重结晶抑制）和进一步的蛋白质活性特征测定方法（冰坑测定法、成核温度测定法）。
