Secondary structure composition of reconstituted subunit b of the Escherichia coli ATP synthase

Autor(en): Greie, JC
Deckers-Hebestreit, G 
Altendorf, K 
Stichwörter: 2ND STALK; ALPHA-SUBUNIT; ATP synthase; Biochemistry & Molecular Biology; CD; CIRCULAR-DICHROISM SPECTROSCOPY; CROSS-LINKING; DELTA-SUBUNIT; DODECYL-SULFATE; EPSILON-SUBUNIT; Escherichia coli; F0F1-ATP SYNTHASE; GRADIENT-INDUCED CHANGES; H+ TRANSLOCATION; secondary structure; subunit b
Erscheinungsdatum: 2000
Herausgeber: BLACKWELL SCIENCE LTD
Journal: EUROPEAN JOURNAL OF BIOCHEMISTRY
Volumen: 267
Ausgabe: 10
Startseite: 3040
Seitenende: 3048
Zusammenfassung: 
Subunit b of the Escherichia coli ATP synthase was isolated by preparative gel electrophoresis, acetone precipitated and after ion-pair extraction redissolved in a buffer either containing n-dodecyl-beta-d-maltoside or sodium cholate. The secondary structure of isolated subunit b was shown to be the same as within the F-O complex, but was strongly dependent on the detergent used for replacement of the phospholipid environment. This was shown by an identical tryptic digestion pattern, which was strongly influenced by the detergent used for solubilization. An influence of the detergent n-dodecyl-beta-d-maltoside on the secondary structure of the hydrophilic part of subunit b was also shown for the soluble part of the polypeptide comprising residues Val25 to Leu156 (b(sol)) using CD spectroscopy. In order to determine the secondary structure of subunit b in its native conformation, isolated subunit b was reconstituted into E. coli lipid vesicles and analyzed with CD spectroscopy. The resulting spectrum revealed a secondary structure composition of 80% alpha helix together with 14% beta turn conformation. These results suggest that subunit b is not a rigid rod-like alpha helix simply linking F-1 to F-O, but rather provides an inherent flexibility for the storage of elastic energy within the second stalk generated by rotational movements within the F1FO complex.
ISSN: 00142956
DOI: 10.1046/j.1432-1033.2000.01327.x

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