A fusion protein of the BRK-DNA binding domain with a FLAG epitope tag was purified [16] and captured onto the anti-Flag antibody.
We have previously demonstrated that RKIP/Raf interaction is regulated by two mechanisms: phosphorylation of RKIP at Ser-153, and occupation of RKIP's conserved ligand binding domain with a phospholipid (2-dihexanoyl-sn-glycero-3-phosphoethanolamine; DHPE).
For bait and prey construction, the PCR products were cloned at EcoR1 sites of pGAD T7 (Pro, P10, P8, RdRp, VPg and CP) vector having the Gal 4 activation domain with a N terminal hemagglutinin (HA) epitope tag and pGBK T7 (MP and MP deletion mutants) vector having Gal 4 DNA binding domain with a N terminal c-Myc epitope tag.
We also found a most interesting fusion of anticodon binding domain with a serine-threonine kinase (PF14_0423) in P. falciparum.
The "RheoReceptor-1" is a fusion of the GAL4 DNA binding domain with a modified ecdysone receptor (EcR) ligand binding domain.
The variable domain of the HCAb (VHH) has a single binding domain with a specificity and affinity similar to conventional antibodies [ 4, 5].
In the other hand, the FMNAT-module consists of an α/β dinucleotide binding domain with a typical Rossmann fold topology (~ 150 aa) [ 9- 11].
Regardless of the level of overall sequence identity, these enzymes share a common conserved S-adenosyl-L-methionine (SAM) binding domain with a core α/ß Rossman fold [ 18] and a unique α-helical cap that forms the top of the active site cavity.
To determine whether POT1 influences the processing of the 3' G-rich single-stranded overhang by WRN, we preincubated POT1 or a mutant form of POT1 (POT1Δ), which lacks the single-stranded DNA binding domain, with a telomeric DNA substrate at room temperature for 20 minutes before the addition of WRN and further incubation at 37C for 15 minutes.
To define the position and number of CENP-A binding domains with a different approach, we designed a set of 27 primer pairs (Supplementary Table 1) spanning the 500 kb region.
