Crystallization and preliminary X-ray crystallographic analysis of subunit F (F1-94), an essential coupling subunit of the eukaryotic V1VO-ATPase from Saccharomyces cerevisiae

• Main Authors: Basak, Sandip, Balakrishna, Asha Manikkoth, Manimekalai, Malathy Sony Subramanian, Grüber, Gerhard
• Other Authors: School of Biological Sciences
• Format: Article
• Language: English
• Published: 2013
• Subjects: DRNTU::Science::Chemistry::Biochemistry; DRNTU::Science::Chemistry::Crystallography
• Online Access: https://hdl.handle.net/10356/95360; http://hdl.handle.net/10220/9180
• Institution: Nanyang Technological University

V-ATPases are very complex multi-subunit enzymes which function as proton-pumping rotary nanomotors. The rotary and coupling subunit F (F1-94) was crystallized by the hanging-drop vapour-diffusion method. The native crystals diffracted to a resolution of 2.64 Å and belonged to space group C2221, with unit-cell parameters a = 47.21, b = 160.26, c = 102.49 Å. The selenomethionyl form of the F1-94 I69M mutant diffracted to a resolution of 2.3 Å and belonged to space group C2221, with unit-cell parameters a = 47.22, b = 160.83, c = 102.74 Å. Initial phasing and model building suggested the presence of four molecules in the asymmetric unit.