title: Probing the recognition properties of the antiparallel coiled coil motif from PKN by protein grafting.
abstract: Coiled coils have long been recognized as the major constituent of many fibrous proteins and also serve as oligomerization domains in a wide variety of proteins. More recently, it has become clear that the surfaces of two-stranded coiled coils are also involved in macromolecular recognition. Indeed, the helical hairpin or intramolecular antiparallel coiled coil (ACC) can serve as a protein or nucleic acid recognition motif. Protein kinase N (PKN) interacts with the small GTPase RhoA through ACC motifs. The crystal structure of RhoA with the N-terminal ACC motif (PKN-ACC1) is unusual in that these proteins interact through two distinct surfaces. Using the ACC domain of seryl tRNA synthetase (SRS-ACC) as a scaffold for protein grafting experiments, we show that RhoA interacts with only one face of PKN-ACC1. This result highlights the potential of the SRS-ACC scaffold for protein engineering applications and provides insight into the mechanism of RhoA-mediated signal transduction through PKN.
authors.affiliation: Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405-7102, USA.
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Li
authors.forename: Harmeet
authors.initials: H
authors.lastname: Kaur
authors.forename: Martha G
authors.initials: MG
authors.lastname: Oakley
mesh_terms: D020816:Amino Acid Motifs; D002942:Circular Dichroism; D003062:Codon; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D008968:Molecular Conformation; D011485:Protein Binding; D011487:Protein Conformation; D011493:Protein Kinase C / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D020349:Surface Plasmon Resonance; D013696:Temperature; D020742:rhoA GTP-Binding Protein / Q000737:chemistry*
chemical_list: D003062:Codon; D000939:Epitopes; C066737:RHOA protein, human; C061570:protein kinase N; D011493:Protein Kinase C; D020742:rhoA GTP-Binding Protein