TY - JOUR
T1 - Isolation of Mitochondria from Ustilago maydis Protoplasts
AU - Pardo, Juan Pablo
AU - Guerra-Sánchez, Guadalupe
AU - Flores-Herrera, Oscar
AU - Romero-Aguilar, Lucero
N1 - Publisher Copyright:
Copyright © 2022 The Authors; exclusive licensee Bio-protocol LLC.
PY - 2022/1/5
Y1 - 2022/1/5
N2 - Ustilago maydis, a basidiomycete that infects Zea mays, is one of the top ten fungal models for studying DNA repair, signal transduction pathways, and dimorphic transitions, among other processes. From a metabolic point of view, U. maydis lacks fermentative capacity, pointing to mitochondria as a key player in central metabolism. Oxidative phosphorylation, synthesis of heme groups, Krebs cycle, β-oxidation of fatty acids, and synthesis of amino acids are some of the processes that take place in mitochondria. Given the importance of this organelle in eukaryotic cells in general, and in fungal cells in particular, we present a protocol for the isolation of U. maydis mitochondria based on the enzymatic disruption of U. maydis cell wall and differential centrifugation. The method can easily be extrapolated to other fungal species, by using appropriate lytic enzymes.
AB - Ustilago maydis, a basidiomycete that infects Zea mays, is one of the top ten fungal models for studying DNA repair, signal transduction pathways, and dimorphic transitions, among other processes. From a metabolic point of view, U. maydis lacks fermentative capacity, pointing to mitochondria as a key player in central metabolism. Oxidative phosphorylation, synthesis of heme groups, Krebs cycle, β-oxidation of fatty acids, and synthesis of amino acids are some of the processes that take place in mitochondria. Given the importance of this organelle in eukaryotic cells in general, and in fungal cells in particular, we present a protocol for the isolation of U. maydis mitochondria based on the enzymatic disruption of U. maydis cell wall and differential centrifugation. The method can easily be extrapolated to other fungal species, by using appropriate lytic enzymes.
KW - Differential centrifugation
KW - Lysing cell wall
KW - Membrane potential
KW - Mitochondria isolation
KW - Respiratory chain
KW - Ustilago maydis
UR - http://www.scopus.com/inward/record.url?scp=85123441628&partnerID=8YFLogxK
U2 - 10.21769/BioProtoc.4277
DO - 10.21769/BioProtoc.4277
M3 - Artículo
C2 - 35118170
AN - SCOPUS:85123441628
SN - 2331-8325
VL - 12
JO - Bio-protocol
JF - Bio-protocol
IS - 1
M1 - e4277
ER -