Motifs involved in interchain binding at the tail-end of spectrin.
Segments 20-22 of alpha-spectrin and 1-3 of beta-spectrin are required for high avidity interchain binding at the tail-end of the molecule. Here, sequence analysis guided by the crystal structure of spectrin's repeating segments was used to redefine the boundaries of a repetitive beta segment that is critical for interchain binding and demonstrate the contribution of non-repetitive spectrin segments in high avidity interchain binding. Our results show that several motifs together are required for high avidity binding, indicating that interchain binding at the tail-end of the spectrin molecule depends on the long distance coordination of several different elements. We also explored the role of unusual motifs contained in beta segments involved in interchain binding. A row of basic residues and a row of small hydrophobic residues were found not to be required for interchain binding, suggesting that their conservation among species reflects functions unrelated to interchain binding. The octamer between segments beta 2 and beta 3 that maintains a specific register between true binding sites was found to have an indirect role in interchain binding by stabilizing neighboring segments. A 5-residue domain in segment beta 2 (EKPPK) was required for interchain binding because it sustains normal helix-helix interactions within segments beta 2.
Viel
A
A
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Gee
M S
MS
Tomooka
L
L
Branton
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D004330:Drosophila / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011487:Protein Conformation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013049:Spectrin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D019476:Insect Proteins; D013049:Spectrin