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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Nat Methods. 2014 Nov 2;11(12):1253–1260. doi: 10.1038/nmeth.3170

Table 1.

Characteristics, affinities, and specificities for LaG, LaM, and LaG dimer proteins. Kds for GFP and mCherry binding were determined by SPR unless otherwise noted. Kds are also shown for LaG dimers fused using a glycine-rich peptide linker (3 repeats of GGGGS, or G4S), or a 3xFLAG linker. For yeast Nup84-GFP and mammalian RBM7-GFP affinity isolations using LaG-conjugated Dynabeads, Coomassie-stained bands from elutions separated by SDS-PAGE were quantified, and known specific and nonspecific bands were used to calculate signal to noise (S:N) ratios. Bead binding assays were used to determine affinity for variant fluorescent proteins, and divergent AmCFP (for LaGs) or DsRed (for LaMs) binding abilities are shown. GFP epitopes were determined by NMR for LaGs, and grouped into three broad classes (I–III). The number of residues identified in the binding site, and its calculated accessible surface area (ASA), are also shown.

Clone ID Mol. Wt (Da) Kd (nM) Nup84-GFP S:N RBM7-GFP S:N Binds AmCFP (LaG)/DsRed (LaM) GFP Epitope No. of binding site residues ASA of binding site residues (Å2)
LaG-2 15,919 191, 16 1.03 0.42 − III 55 2,204
LaG-3 15,329 25 0.77 1.13 + nd nd nd
LaG-6 15,700 310 0.12 nd + nd nd nd
LaG-9 16,062 3.5 1.02 1.04 + I 62 2,551
LaG-10 15,748 97 0.17 nd + nd nd nd
LaG-12 16,090 56 0.20 nd + nd nd nd
LaG-14 16,002 1.9 0.84 0.58 + I 66 2,519
LaG-16 16,306 0.7 1.05 0.92 + I 60 2,605
LaG-17 15,823 50 0.67 nd + I 60 2,543
LaG-19 15,528 24.61 0.95 1.06 + II 54 2,404
LaG-21 15,452 7 1.09 nd + II 56 2,340
LaG-24 14,763 41 1.05 1.09 − III2 nd nd
LaG-26 16,221 2.6 1.00 nd + II 53 2,070
LaG-27 15,565 9.5 1.04 nd + II 57 2,216
LaG-29 15,449 110 0.31 nd + nd nd nd
LaG-30 16,159 0.5 1.04 nd + nd nd nd
LaG-35 16,010 23.51 0.70 nd + nd nd nd
LaG-37 16,329 24 0.36 nd + nd nd nd
LaG-41 15,471 0.9 1.12 0.41 + II 53 2,091
LaG-42 15,490 600 0.21 nd + nd nd nd
LaG-43 16,167 11 0.69 nd + I 55 2,381
LaG-5 15,589 14,2001 0.11 nd nd nd nd nd
LaG-8 15,953 20,0001 0.10 nd nd nd nd nd
LaG-11 16,221 22,9001 0.10 nd nd nd nd nd
LaG-18 16,459 3,8001 0.13 nd nd nd nd nd
LaG16-G4S-2 30,791 0.036 nd nd nd nd nd nd
LaG16-3xFLAG-2 32,972 0.268 nd nd nd nd nd nd
LaG41-G4S-2 29,956 0.150 nd nd nd nd nd nd
LaM-1 15,380 22 n/a n/a − n/a n/a n/a
LaM-2 15,151 0.49 n/a n/a − n/a n/a n/a
LaM-3 15,196 1.9 n/a n/a + n/a n/a n/a
LaM-4 14,866 0.18 n/a n/a + n/a n/a n/a
LaM-6 14,428 0.26 n/a n/a − n/a n/a n/a
LaM-8 14,666 63 n/a n/a − n/a n/a n/a
1

Kd determined by bead binding assay.

2

Determined by binding assays and mutagenesis.