Journal: Molecular Biology and Evolution

Article Title: The Core MICOS Complex Subunit mic60 has Been Substituted by Two Cryptic Mitofilin-containing Proteins in Euglenozoa

doi: 10.1093/molbev/msaf289

Figure Lengend Snippet: Mic34 binds to phospholipid bilayers in vitro. a) Carbonate extraction of Mic34 loaded LUVs with 25 or 75 mM sodium carbonate. Supernatant, S; pellet, P. b) Proteinase K (PK) protection assay of LUVs loaded with Mic34 or Mic19 and Folch liposomes loaded with ENTH visualized with Coomassie Brilliant Blue staining after SDS-PAGE. c) Flotated LUVs with and without (-CL) cardiolipin separated on aa noncontinuous Histodenz density gradient (schema on left) loaded with recombinant Mic34. Compare banding pattern with LUVs flotated alone (right). d) Histodenz density gradient fractions resolved by SDS-PAGE and stained with Coomassie Brilliant Blue or transferred to a membrane for incubation with an anti-His antibody (Mic34 with and without CL). e) Negatively stained LUVs loaded with Mic34. Scale bar, 100 nm. f) Negatively stained Folch + 5% PI(4,5)P2 liposomes incubated with the membrane remodeling protein ENTH that did (+DDM) or did not (DDM-) undergo prior solubilization with 0.1% DDM. Scale bar, 200 nm.

Article Snippet: Egg l-α-phosphatidylcholine (PC), egg l-α-phosphatidylethanolamine (PE), liver l-α-phosphatidylinositol (PI), brain l-α-phosphatidylserine (PS), Heart cardiolipin (CL), and brain L-α-phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) were purchased from Avanti Polar Lipids, Inc. Folch bovine brain extract Fraction I was purchased from Sigma Aldrich.

Techniques: In Vitro, Extraction, Liposomes, Staining, SDS Page, Recombinant, Membrane, Incubation