The repressor of primer (Rop) protein has become a steady source of surprises concerning the relationship between the sequences and the structures of several of its mutants and variants. Here we add another piece to the puzzle of Rop by showing that an engineered deletion mutant of the protein (corresponding to a deletion of residues 30-34 of the wild-type protein and designed to restore the heptad periodicity at the turn region) results in a complete reorganization of the bundle which is converted from a homodimer to a homotetramer. In contrast (and as previously shown), a two-residue insertion, which also restores the heptad periodicity, is essentially identical with wild-type Rop. The new deletion mutant structure is a canonical, left-handed, all-antiparallel bundle with a completely different hydrophobic core and distinct surface properties. The structure agrees and qualitatively explains the results from functional, thermodynamic, and kinetic studies which indicated that this deletion mutant is a biologically inactive hyperstable homotetramer. Additional insight into the stability and dynamics of the mutant structure has been obtained from extensive molecular dynamics simulations in explicit water and with full treatment of electrostatics.

Glykos, N., Papanikolau, Y., Vlassi, M., Kotsifaki, D., Cesareni, G., Kokkinidis, M. (2006). Loopless rop: Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein. BIOCHEMISTRY, 45(36), 10905-10919 [10.1021/bi060833n].

Loopless rop: Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein

CESARENI, GIOVANNI;
2006-01-01

Abstract

The repressor of primer (Rop) protein has become a steady source of surprises concerning the relationship between the sequences and the structures of several of its mutants and variants. Here we add another piece to the puzzle of Rop by showing that an engineered deletion mutant of the protein (corresponding to a deletion of residues 30-34 of the wild-type protein and designed to restore the heptad periodicity at the turn region) results in a complete reorganization of the bundle which is converted from a homodimer to a homotetramer. In contrast (and as previously shown), a two-residue insertion, which also restores the heptad periodicity, is essentially identical with wild-type Rop. The new deletion mutant structure is a canonical, left-handed, all-antiparallel bundle with a completely different hydrophobic core and distinct surface properties. The structure agrees and qualitatively explains the results from functional, thermodynamic, and kinetic studies which indicated that this deletion mutant is a biologically inactive hyperstable homotetramer. Additional insight into the stability and dynamics of the mutant structure has been obtained from extensive molecular dynamics simulations in explicit water and with full treatment of electrostatics.
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/18 - Genetica
English
Con Impact Factor ISI
Dimers; Electrostatics; Molecular dynamics; Mutagenesis; Surface properties; Thermodynamics; Homodimers; Homotetramers; Mutants; Primer protein; Proteins; homodimer; homotetramer; mutant protein; repressor protein; Rop protein; tetramer; unclassified drug; water; article; deletion mutant; electricity; enzyme structure; molecular dynamics; priority journal; protein engineering; protein function; protein stability; protein variant; sequence analysis; simulation; surface property; thermodynamics; wild type; Amino Acid Sequence; Bacterial Proteins; Crystallography, X-Ray; Hydrophobicity; Models, Molecular; Molecular Sequence Data; Protein Conformation; Recombinant Proteins; Repetitive Sequences, Amino Acid; RNA-Binding Proteins
Glykos, N., Papanikolau, Y., Vlassi, M., Kotsifaki, D., Cesareni, G., Kokkinidis, M. (2006). Loopless rop: Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein. BIOCHEMISTRY, 45(36), 10905-10919 [10.1021/bi060833n].
Glykos, N; Papanikolau, Y; Vlassi, M; Kotsifaki, D; Cesareni, G; Kokkinidis, M
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/35795
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