Multiple sequence alignment of NDE1 and NDEL1 orthologs.
Orthologs of NDE1 and NDEL1 across vertebrate species were identified using the UCSC Genome Browser (http://genome-euro.ucsc.edu). These were aligned initially using Clustal Omega (108, 109) with further manual editing for optimum alignment. The sequence numbering at the top of each alignment block corresponds to the sequence of human NDE1. Conserved residues are colored with a red background; conservatively substituted residues are shown with a yellow background. The location of the N-terminal coiled-coil α-helical domain (residues ∼10–185) and the predicted C-terminal α-helix (residues ∼247–278) are shown above the alignment. Consensus amino acids are shown at the bottom of the each alignment block (uppercase is strictly conserved; lowercase is consensus level >0.5; ! is I or V; $ is L or M; % is F or Y; = is N, D, Q or E). Despite 60% sequence identity there are at least 18 specific phosphorylation sites for each human protein – i.e., phosphorylation possible only in one protein. Some of these phosphorylation sites have been derived from high-throughput proteomic screening using mass spectrometry (90), while others have been experimentally verified in the cell. See the main text and for more information. Those sites known to be specific to either NDE1 or NDEL1 are highlighted with a green filled oval under the corresponding star; black star=phosphorylation site in NDE1; orange star=phosphorylation site in NDEL1; pink star=phosphorylation site in both NDE1 and NDEL1; cyan block=palmitoylation site in both proteins. The figure was generated with ESPript v2.2 (111).