Chemical Probe for Imaging of Polo-like Kinase 4 and Centrioles

Authors:

Aleksandar Salim, Philipp Werther, Georgios N. Hatzopoulos, Luc Reymond, Richard Wombacher, Pierre Gönczy, and Kai Johnsson

Keywords:

Polo-like kinase, Plk-4, Cen-TCO, centriole, protein labeling, bio-orthogonal chemistry, fluorogenic fluorophores

Abstract:

Polo-like kinase (Plk4) is a serine/threonine-protein kinase that is essential for biogenesis of the centriole organelle and is enriched at centrioles. Herein, we introduce Cen-TCO, a chemical probe based on the Plk4 inhibitor centrinone, to image Plk4 and centrioles in live or fixed cultured human cells. Specifically, we established a bio-orthogonal two-step labeling system that enables the Cen-TCO-mediated imaging of Plk4 by STED super-resolution microscopy. Such direct labeling of Plk4 results in an increased resolution in STED imaging compared with using anti-Plk4 antibodies, underlining the importance of direct labeling strategies for super-resolution microscopy. We anticipate that Cen-TCO will become an important tool for investigating the biology of Plk4 and of centrioles.

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