Structural and functional analysis of the coupling subunit F in solution and topological arrangement of the stalk domains of the methanogenic A1AO ATP synthase.
The first low-resolution shape of subunit F of the A(1)A(O) ATP synthase from the archaeon Methanosarcina mazei Gö1 in solution was determined by small angle X-ray scattering. Independent to the concentration used, the protein is monomeric and has an elongated shape, divided in a main globular part with a length of about 4.5 nm, and a hook-like domain of about 3.0 nm in length. The subunit-subunit interaction of subunit F inside the A(1)A(O) ATP synthase in the presence of 1-ethyl-3-(dimethylaminopropyl)-carbodiimide EDC was studied as a function of nucleotide binding, demonstrating movements of subunits F relative to the nucleotide-binding subunit B. Furthermore, in the intact A(1)A(O) complex, crosslinking of subunits D-E, A-H and A-B-D was obtained and the peptides, involved, were analyzed by MALDI-TOF mass spectrometry. Based on these data the surface of contact of B-F could be mapped in the high-resolution structure of subunit B of the A(1)A(O) ATP synthase.
Schäfer
Ingmar
I
School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.
Rössle
Manfred
M
Biuković
Goran
G
Müller
Volker
V
Grüber
Gerhard
G
D025181:ATP Synthetase Complexes / Q000737:chemistry*; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives / Q000737:chemistry; D017020:Methanosarcina / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry
D019843:Archaeal Proteins; D003432:Cross-Linking Reagents; D021122:Protein Subunits; C041829:1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide methiodide; D025181:ATP Synthetase Complexes; D005022:Ethyldimethylaminopropyl Carbodiimide