Putative HspB2 binding partners retrieved from the Y2H screen reveal a role in muscle and mitochondrial maintenance.

* Indicates the 30 proteins shown to be HspB2 dependent in the Y2H binding assay. All others were not tested for dependency. Of the total 49 plasmids tested for dependency from the multi-hitter list, 10 did not appear in frame from sequence analysis, 81.6% showed a positive Y2H result that was dependent on HspB2 (indicating a HspB2/prey interaction), 10.2% were false positives (they grew with either HspB2 or the empty bait plasmid), and 8.2% were true negatives (they did not grow with either HspB2 or the empty bait). Out of frame fusions, false positives and true negatives revealed through this analysis are not listed.** Abbreviations include Endoplasmic Reticulum (ER), Mitochondrial (Mito), Mitochondrial Inner Membrane (Mito IM), Mitochondrial Outer Membrane (Mito OM), Mitochondrial Matrix (Mito Matrix), Plasma Membrane (PM) and Sarcoplasmic Reticulum (SR).]Indicates that the final amino acid listed is the C-terminal amino acid of the native protein.Indicates that there is extra DNA from the mitochondrial chromosome on the 3’ end of this construct in addition to the MYH6 DNA which adds an extra 82 aa.# Indicates that the sequence quality was too poor to unambiguously tell the N-terminal fusion (WDR1, NHP2L, ATP6V1E1, TATDN1, MLF2) or the C-terminal end (RYR2, WDR1, ALDH2, ENO1, HADHB, MDH1, PGM1, AGAP11, MYOZ2, ANF335, GARS, CKM, ABCC9, EEF1A1, THUMPD2, A2M).Proteins retrieved multiple times from a Y2H screen for HspB2 binding partners (84 hits) are given along with the following information: protein name, UniProt [73, 74] protein accession number (hyperlink), gene symbol, primary cellular localization reported on HPRD [76] or UniProt, the number of times it was retrieved from the screen (# hits) and the amino acids corresponding to the library construct from the n-terminal amino acid to the c-terminal amino acid (if the terminal amino acid is not reached the total amino acids are given in parenthesis). In some cases the nucleotide fusion occurred before the initiating ATG and is indicated by the number of amino acids upstream of the start (for example, -12–136 for LGALS1 denotes 22 amino acids upstream of the ATG start to amino acid 136). Hits that did not correspond to a coding sequence are not listed and include the 3’ region of ABLIM1, AXIN1, DYRK1A, FHL1, GPX3, HIPK2, NCAM1, PPAPDC3, PLXNA4, LRP10, SCN1B, SEC62, SGSM2, SLC25A4, SNTA1, SYNPO, TNIP1, USP28, and XIRP1 mRNA. The proteins are functionally categorized by their description on UniProt [73, 74] and/or GeneCards [73]. See supplementary S1 Table for a complete list of putative HspB2 binding partners retrieved from the Y2H screen only once.