Table 1.

Affinity purification of the ERMES complex identifies an uncharacterized protein, Ylr072w

Identified protein Average spectral counts Description CRAPome score 
 Mdm12 Mdm34 Mmm1   
Mmm1 13 30 71 ERMES subunit 0.00 
Mdm34 17 48 37 ERMES subunit 0.00 
Gem1 12 35 34 Miro GTPase 0.00 
Ylr072w 17.5 5.5 Uncharacterized 0.00 
Mdm12 ERMES subunit 0.00 
Msc7 ER protein 0.00 
Tom71 7.5 Mitochondrial protein import receptor 0.00 
Pbn1 Component of GPI-mannosyltransferase I 0.00 
Pho84 1.5 4.5 Abundant inorganic phosphate transporter 0.00 
Vtc1 Vacuolar transporter complex 0.00 
Spt10 1.5 Histone acetylase 0.06 
Mdm10 26 20.5 ERMES subunit 0.12 
Myo2 Type V myosin; mitochondria/vacuole transport 0.18 
Lsp1 2.5 Eisosome protein 0.18 
Por1 6.5 22.5 17.5 Mitochondrial porin 0.35 
Fks1 0.5 β-Glucan synthase component 0.41 
Identified protein Average spectral counts Description CRAPome score 
 Mdm12 Mdm34 Mmm1   
Mmm1 13 30 71 ERMES subunit 0.00 
Mdm34 17 48 37 ERMES subunit 0.00 
Gem1 12 35 34 Miro GTPase 0.00 
Ylr072w 17.5 5.5 Uncharacterized 0.00 
Mdm12 ERMES subunit 0.00 
Msc7 ER protein 0.00 
Tom71 7.5 Mitochondrial protein import receptor 0.00 
Pbn1 Component of GPI-mannosyltransferase I 0.00 
Pho84 1.5 4.5 Abundant inorganic phosphate transporter 0.00 
Vtc1 Vacuolar transporter complex 0.00 
Spt10 1.5 Histone acetylase 0.06 
Mdm10 26 20.5 ERMES subunit 0.12 
Myo2 Type V myosin; mitochondria/vacuole transport 0.18 
Lsp1 2.5 Eisosome protein 0.18 
Por1 6.5 22.5 17.5 Mitochondrial porin 0.35 
Fks1 0.5 β-Glucan synthase component 0.41 

Functional GFP fusions to the ERMES proteins Mdm12, Mdm34, and Mmm1 encoded at their endogenous chromosomal loci were purified from yeast cell lysates with GFP antibodies. Purified proteins were identified by tandem mass spectrometry with a <1% decoy false discovery rate. As a control, mock purifications were performed from an isogenic strain background that did not express GFP. The average total spectrum counts assigned to each protein from two IPs for Mdm12 and Mmm1 and four IPs for Mdm34 are tabulated. We included in this table proteins that met the following criteria: (a) they were identified in duplicate sample experiments for Mdm12/Mmm1 or 3/4 IPs for Mdm34, and (b) their average spectral counts were at least 10-fold higher than in mock purifications. Tabulated are proteins that were identified in duplicate sample experiments and whose average spectral counts were at least 10-fold higher than in mock purifications. The “CRAPome” score for identified proteins is listed at right and is a measure of how likely the identified protein is the result of a nonspecific interaction. This number is derived from the Contaminant Repository for Affinity Purifications database, CRAPome, and is the proportion of mock purifications in which the protein is identified.

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