ABCMdb

PMID: 26195633

Camire EJ, Grossman JD, Thole GJ, Fleischman NM, Perlstein DL
The Yeast Nbp35-Cfd1 Cytosolic Iron-Sulfur Cluster Scaffold Is an ATPase.
J Biol Chem. 2015 Sep 25;290(39):23793-802. doi: 10.1074/jbc.M115.667022. Epub 2015 Jul 20., [PubMed]

Sentences

No. Mutations Sentence Comment

138 ABCC1 p.Lys31Ala Lane 1, His Nbp35; lane 2, His Cfd1; lane 3, Nbp35-Cfd1 complex; lane 4, Nbp35-K26A Cfd1; lane 5, Nbp35-K31A Cfd1; lane 6, K86A Nbp35-Cfd1. Login to comment 150 ABCC1 p.Lys31Ala The lysine mutants K86A Nbp35-Cfd1, Nbp35-K26A Cfd1, and Nbp35-K31A Cfd1 were expressed at similar levels as the wild-type Nbp35-Cfd1 complex, whereas little K81A Nbp35 was present in crude extracts upon co-expression with wild-type Cfd1. Login to comment 151 ABCC1 p.Lys31Ala For the remaining three mutants, intact complexes containing K86A Nbp35-Cfd1, Nbp35-K26A Cfd1, and Nbp35-K31A Cfd1 could be obtained. Login to comment 154 ABCC1 p.Lys31Ala To confirm via a second independent method that our mutations did not significantly affect the structure of the heterocomplex, we analyzed the secondary structure content of the two canonical lysine mutants, K86A Nbp35-Cfd1 and Nbp35-K31A Cfd1, via CD. Login to comment 202 ABCC1 p.Lys31Ala C shows wild-type Nbp35-Cfd1 complex (afb;), Nbp35-K26A Cfd1 (); Nbp35-K31A Cfd1(Éa;), and Nbp35-D55A Cfd1 (E). Login to comment 220 ABCC1 p.Lys31Ala We found that mutation of either lysine in the Walker A motif, K26A or K31A, resulted in decreased ATP hydrolysis activity of the Nbp35-Cfd1 complex (Fig. 5C, triangles). Login to comment