A dual binding mode for RhoGTPases in plexin signalling by Bell, C, Aricescu, A, Jones, E, Siebold, C

“…On inclusion of the juxtamembrane helix, a trimeric structure of Plexin-B1-Rac1 complexes is stabilised by a second, novel, RhoGTPase binding site adjacent to the Ras site. Site-directed mutagenesis combined with cellular and biophysical assays demonstrate that this new binding site is essential for signalling. …”

Mutations in alpha-tubulin cause abnormal neuronal migration in mice and lissencephaly in humans. by Keays, D, Tian, G, Poirier, K, Huang, G, Siebold, C, Cleak, J, Oliver, P, Fray, M, Harvey, R, Molnár, Z, Piñon, M, Dear, N, Valdar, W, Brown, S, Davies, K, Rawlins, J, Cowan, N, Nolan, P, Chelly, J, Flint, J

“…This study demonstrates the utility of ENU mutagenesis in the mouse as a means to discover the basis of human neurodevelopmental disorders.…”

Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. by Bishop, B, Aricescu, A, Harlos, K, O'Callaghan, C, Jones, E, Siebold, C

“…The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. …”

Structure and functional analysis of the IGF-II/IGF2R interaction. by Brown, J, Delaine, C, Zaccheo, O, Siebold, C, Gilbert, R, van Boxel, G, Denley, A, Wallace, J, Hassan, A, Forbes, B, Jones, E

“…Our complex shows that Phe19 and Leu53 of IGF-II lock into a hydrophobic pocket unique to domain 11 of mammalian IGF2Rs. Mutagenesis analyses confirm this IGF-II 'binding-hotspot', revealing that IGF-binding proteins and IGF2R have converged on the same high-affinity site.…”

R-spondins engage heparan sulfate proteoglycans to potentiate WNT signaling by Dubey, R, van Kerkhof, P, Jordens, I, Malinauskas, T, Pusapati, GV, McKenna, JK, Li, D, Carette, JE, Ho, M, Siebold, C, Maurice, M, Lebensohn, AM, Rohatgi, R

“…We now show that heparan sulfate proteoglycans (HSPGs) act as alternative co-receptors for RSPO3 using a combination of ligand mutagenesis and ligand engineering. Mutations in RSPO3 residues predicted to contact HSPGs impair its signaling capacity. …”

A dual binding mode for RhoGTPases in plexin signalling by Bell, C, Aricescu, A, Jones, E, Siebold, C

“…On inclusion of the juxtamembrane helix, a trimeric structure of Plexin-B1-Rac1 complexes is stabilised by a second, novel, RhoGTPase binding site adjacent to the Ras site. Site-directed mutagenesis combined with cellular and biophysical assays demonstrate that this new binding site is essential for signalling. …”

Structural insights into proteoglycan-shaped Hedgehog signaling. by Whalen, D, Malinauskas, T, Gilbert, R, Siebold, C

“…The interaction determinants, confirmed by site-directed mutagenesis and binding studies, reveal a previously not identified Hh site for GAG binding, common to all Hh proteins. …”

Molecular analysis of receptor protein tyrosine phosphatase mu-mediated cell adhesion. by Aricescu, A, Hon, W, Siebold, C, Lu, W, Van Der Merwe, P, Jones, E

“…Structure-based site-directed mutagenesis, serial domain deletions and cell-adhesion assays allowed us to identify the four N-terminal domains (MAM, Ig, fibronectin type III (FNIII)-1 and FNIII-2) as a minimal functional unit. …”

Molecular mechanisms of repulsive guidance molecule signalling by Healey, E

“…In Chapter 3, an analysis of an RGMB-NEO1 crystal structure is presented. Combined with mutagenesis studies, analytical ultracentrifugation experiments and neurite outgrowth assays, this allowed a mechanism for RGM signalling through NEO1 to be proposed. …”

A calcium-sensing receptor mutation causing hypocalcemia disrupts a transmembrane salt bridge to activate β-arrestin-biased signaling by Gorvin, C, Babinsky, V, Malinauskas, T, Nissen, P, Schou, A, Hanyaloglu, A, Siebold, C, Jones, E, Hannan, F, Thakker, R

“…Homology modeling and mutagenesis studies showed that the R680G CaSR mutation selectively enhanced β-arrestin signaling by disrupting a salt bridge formed between Arg680 and Glu767, which are located in CaSR transmembrane domain 3 and extracellular loop 2, respectively. …”

Structural and functional studies of the hedgehog signalling pathway by Whalen, D

“…The interaction determinants, confirmed by binding studies and mutagenesis reveal a novel Hh site for GAG interactions, which appears to be common to all Hh proteins. …”

Hedgehog Acyltransferase: Structure, Function and Inhibition by Coupland, C

“…Furthermore, structural information is confirmed through mutagenesis assays. In Chapter 6, the interaction of HHAT with the acyl-receiving substrate, SHH, is explored through the use of biophysical approaches to identify the SHH binding site. …”