Characterization of the tetratricopeptide-containing domain of BUB1, BUBR1, and PP5 proves that domain amphiphilicity over amino acid sequence specificity governs protein adsorption and interfacial activity.
The tetratricopeptide motif repeat (TPR) is an alpha-helix-turn-alpha-helix motif that typically mediates protein-protein and, in some cases, protein-lipid interactions. Because of its success, this motif has been preserved through evolution and can be identified in proteins of a wide range of functions in lower and higher organisms. The N-terminal region of BUB1, BUBR1, and protein phosphatase 5 (PP5) contains tandem arrangements of the TPR motif. BUB1 and BUBR1 are conserved multidomain protein kinases that play a key role in the mitotic checkpoint, the mechanism that ensures the synchrony of chromosome segregation. PP5 is an enzyme that targets a wide range of protein substrates including single transmembrane receptors and mammalian cryptochromes. The N-terminal TPR domain of PP5 regulates the activity of the C-terminal catalytic domain through direct interaction with protein and lipid molecules. We portray the biophysical and biochemical properties of the tandem arrangements of the TPR motif of BUB1, BUBR1, and PP5 using far-UV spectroscopy, solution X-ray scattering, null ellipsometry, surface rheology measurements, and Brewster angle microscopy (BAM) observations. We show that, despite the low amino acid sequence conservation and different function, the TPR motif repeats of the three proteins exhibit similar interfacial properties including adsorption kinetics, high surface activity, and the formation of stable, rigid films at the air/water interface. Our studies demonstrate that domain amphiphilicity is of higher importance than amino acid sequence specificity in the determination of protein adsorption and interfacial activity.
Department of Biochemistry, University of Cambridge, Cambridge, UK.
Sylvie
S
Beaufils
J Günter
JG
Grossmann
Anne
A
Renault
Victor M
VM
Bolanos-Garcia
D000327:Adsorption; D000388:Air; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D016718:Arachidonic Acid / Q000378:metabolism; D004548:Elasticity; D004795:Enzyme Stability; D019143:Evolution, Molecular; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism*; D012212:Rheology; D013379:Substrate Specificity; D013499:Surface Properties; D014867:Water / Q000737:chemistry
D009687:Nuclear Proteins; D014867:Water; D016718:Arachidonic Acid; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases; D010749:Phosphoprotein Phosphatases; C089641:protein phosphatase 5