Figure S4. The C-terminal ends of 220... | Rockefeller University Press

Figure S4.

$The C-terminal ends of 220 kD/440 kD AnkB each contain another type of intrinsically disordered repeating sequence but does not bind to MTs.(A) Top: Schematic of gAnkB domains including membrane binding domain (MBD), SBD, NSD, and CD (highlighted in red). Bottom left: Coomassie blue–stained SDS-PAGE gel of a MT cosedimentation experiment performed with 2 mg/ml Taxol-polymerized tubulins and indicated recombinant proteins, each at 10 µM (Trx at 30 µM). Supernatant and pellet fractions are labeled as S and P, respectively. The bands of tubulin (∼55 kD) are marked by an arrow. CD, gAnkB-F2, and Trx bands are indicated with red boxes. Bottom right: The pellet fractions were quantified on the basis of band intensity and SDS-PAGE sample loading ratio (mean ± SEM; n = 3). (B) Top: Schematic of rat gAnkG fragment containing one 12-aa repeat and a BG-Box. Bottom left: Coomassie blue–stained SDS-PAGE gel of a MT cosedimentation experiment performed with 2 mg/ml prepolymerized tubulins and gAnkG fragment or the Trx control, each at 10 µM (Trx at 30 µM). Bottom right: The pellet fractions were quantified on the basis of band intensity and SDS-PAGE sample loading ratio (mean ± SEM; n = 3).$

The C-terminal ends of 220 kD/440 kD AnkB each contain another type of intrinsically disordered repeating sequence but does not bind to MTs. (A) Top: Schematic of gAnkB domains including membrane binding domain (MBD), SBD, NSD, and CD (highlighted in red). Bottom left: Coomassie blue–stained SDS-PAGE gel of a MT cosedimentation experiment performed with 2 mg/ml Taxol-polymerized tubulins and indicated recombinant proteins, each at 10 µM (Trx at 30 µM). Supernatant and pellet fractions are labeled as S and P, respectively. The bands of tubulin (∼55 kD) are marked by an arrow. CD, gAnkB-F2, and Trx bands are indicated with red boxes. Bottom right: The pellet fractions were quantified on the basis of band intensity and SDS-PAGE sample loading ratio (mean ± SEM; n = 3). (B) Top: Schematic of rat gAnkG fragment containing one 12-aa repeat and a BG-Box. Bottom left: Coomassie blue–stained SDS-PAGE gel of a MT cosedimentation experiment performed with 2 mg/ml prepolymerized tubulins and gAnkG fragment or the Trx control, each at 10 µM (Trx at 30 µM). Bottom right: The pellet fractions were quantified on the basis of band intensity and SDS-PAGE sample loading ratio (mean ± SEM; n = 3).

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