4RGD

The structure a AS-48 G13K/L40K mutant

4RGD の概要

• エントリーDOI: 10.2210/pdb4rgd/pdb
• 関連するPDBエントリー: 1O82 1O83 1O84
• 分子名称: Bacteriocin AS-48, CITRIC ACID (3 entities in total)
• 機能のキーワード: circular bacteriocin, membrane interaction, plasma membrane, antibiotic
• 由来する生物種: Enterococcus faecalis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 14723.50

構造登録者

Sanchez-Barrena, M.J. (登録日: 2014-09-29, 公開日: 2015-04-15, 最終更新日: 2024-11-20)

主引用文献

Cebrian, R.,Martinez-Bueno, M.,Valdivia, E.,Albert, A.,Maqueda, M.,Sanchez-Barrena, M.J.
The bacteriocin AS-48 requires dimer dissociation followed by hydrophobic interactions with the membrane for antibacterial activity.
J.Struct.Biol., 190:162-172, 2015

PubMed Abstract: The molecular mechanism underlining the antibacterial activity of the bacteriocin AS-48 is not known, and two different and opposite alternatives have been proposed. Available data suggested that the interaction of positively charged amino acids of AS-48 with the membrane would produce membrane destabilization and disruption. Alternatively, it has been proposed that AS-48 activity could rely on the effective insertion of the bacteriocin into the membrane. The biological and structural properties of the AS-48G13K/L40K double mutant were investigated to shed light on this subject. Compared with the wild type, the mutant protein suffered an important reduction in the antibacterial activity. Biochemical and structural studies of AS-48G13K/L40K mutant suggest the basis of its decreased antimicrobial activity. Lipid cosedimentation assays showed that the membrane affinity of AS-48G13K/L40K is 12-fold lower than that observed for the wild type. L40K mutation is responsible for this reduced membrane affinity and thus, hydrophobic interactions are involved in membrane association. Furthermore, the high-resolution crystal structure of AS-48G13K/L40K, together with the study of its dimeric character in solution showed that G13K stabilizes the inactive water-soluble dimer, which displays a reduced dipole moment. Our data suggest that the cumulative effect of these three affected properties reduces AS-48 activity, and point out that the bactericidal effect is achieved by the electrostatically driven approach of the inactive water-soluble dimer towards the membrane, followed by the dissociation and insertion of the protein into the lipid bilayer.

PubMed: 25816760
DOI: 10.1016/j.jsb.2015.03.006

実験手法

X-RAY DIFFRACTION (1.2 Å)