ANKRD54 preferentially selects Bruton’s Tyrosine Kinase (BTK) from a Human Src-Homology 3 (SH3) domain library

Abstract

Bruton’s Tyrosine Kinase (BTK) is a cytoplasmic protein tyrosine kinase with a fundamental role in B-lymphocyte development and activation. The nucleocytoplasmic shuttling of BTK is specifically modulated by the Ankyrin Repeat Domain 54 (ANKRD54) protein and the interaction is known to be exclusively SH3-dependent. To identify the spectrum of the ANKRD54 SH3-interactome, we applied phage-display screening of a library containing all the 296 human SH3 domains. The BTK-SH3 domain was the prime interactor. Quantitative western blotting analysis demonstrated the accuracy of the screening procedure. Revealing the spectrum and specificity of ANKRD54-interactome is a critical step toward functional analysis in cells and tissues.

The role of Bruton's tyrosine kinase in the immune system and disease

“…Lyn‐interacting ankyrin repeat protein (LIAR) has been suggested to regulate BTK trafficking via BTK phosphorylation [21]. More recently, Gustafsson et al [22] reported that the ANKRD54 modulates the shuttling of BTK to the nucleus.…”

Generating FN3‐Based Affinity Reagents Through Phage Display

“…Because the DNA sequences of isolated, recombinant antibodies are easily obtained, reagents can be expressed in Escherichia coli , and their affinity and specificity can be improved through directed mutagenesis to yield high‐quality, specific affinity reagents. By this approach, affinity reagents have successfully been generated to recognize a wide range of targets, including cell signaling proteins, membrane proteins, transcription factors, peptides, and post‐translational modifications (Gustafsson et al., ; Hornsby et al., ; Jones et al., ; Kim, Stroud, & Craik, ; Koide & Huang, ; Kummer et al., ; Pershad, Wypisniak, & Kay, ).…”