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

Nicholas M. Glykos, Yannis Papanikolau, Metaxia Vlassi, Dina Kotsifaki, Giovanni Cesareni, Michael Kokkinidis

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)10905-10919
Number of pages15
JournalBiochemistry
Volume45
Issue number36
DOIs
Publication statusPublished - Sep 12 2006

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Repressor Proteins
Periodicity
Surface Properties
Mutant Proteins
Molecular Dynamics Simulation
Static Electricity
Thermodynamics
Surface properties
Molecular dynamics
Electrostatics
Proteins
Kinetics
Water
Computer simulation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Glykos, N. M., 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. https://doi.org/10.1021/bi060833n

Loopless rop : Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein. / Glykos, Nicholas M.; Papanikolau, Yannis; Vlassi, Metaxia; Kotsifaki, Dina; Cesareni, Giovanni; Kokkinidis, Michael.

In: Biochemistry, Vol. 45, No. 36, 12.09.2006, p. 10905-10919.

Research output: Contribution to journalArticle

Glykos, NM, 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, vol. 45, no. 36, pp. 10905-10919. https://doi.org/10.1021/bi060833n
Glykos NM, Papanikolau Y, Vlassi M, Kotsifaki D, Cesareni G, Kokkinidis M. Loopless rop: Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein. Biochemistry. 2006 Sep 12;45(36):10905-10919. https://doi.org/10.1021/bi060833n
Glykos, Nicholas M. ; Papanikolau, Yannis ; Vlassi, Metaxia ; Kotsifaki, Dina ; Cesareni, Giovanni ; Kokkinidis, Michael. / Loopless rop : Structure and dynamics of an engineered homotetrameric variant of the repressor of primer protein. In: Biochemistry. 2006 ; Vol. 45, No. 36. pp. 10905-10919.
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