A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae

被引:3
作者
Cuevas-Bermudez, Abel [1 ]
Garrido-Godino, Ana, I [1 ]
Gutierrez-Santiago, Francisco [1 ]
Navarro, Francisco [1 ,2 ]
机构
[1] Univ Jaen, Dept Biol Expt Genet, Jaen, Spain
[2] Univ Jaen, Ctr Estudios Avanzados Aceite Oliva & Olivar, Paraje Las Lagunillas S-N, Jaen 23071, Spain
来源
BIO-PROTOCOL | 2020年 / 10卷 / 01期
关键词
Chromatin; Proteins; Yeast; Mass spectrometry; Saccharomyces cerevisiae; DNA-REPLICATION; CELL-CYCLE; PROTEINS; PROTEOME;
D O I
10.21769/BioProtoc.3471
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using zymolyase to obtain the spheroplast, which is employed in many other procedures. Employing small amount of culture cells and small volumes of solutions during the yChEFs procedure is very useful to allow many samples to be handled at the same time, and also reduces costs and efforts. The purified proteins associated with chromatin fractions obtained by yChEFs can be analyzed by Western blot (Figure 1) or combined with mass spectrometry for proteomic analyses.
引用
收藏
页数:9
相关论文
共 50 条
  • [31] Purification and characterization of α-keto amide reductase from Saccharomyces cerevisiae
    Ishihara, K
    Yamamoto, H
    Mitsuhashi, K
    Nishikawa, K
    Tsuboi, S
    Tsujl, H
    Nakajima, N
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2004, 68 (11) : 2306 - 2312
  • [32] Purification and characterization of two enone reductases from Saccharomyces cerevisiae
    Wanner, P
    Tressl, R
    EUROPEAN JOURNAL OF BIOCHEMISTRY, 1998, 255 (01): : 271 - 278
  • [33] Purification and characterization of a soluble polyphosphatase from mitochondria of Saccharomyces cerevisiae
    Lichko, LP
    Kulakovskaya, TV
    Kulaev, IS
    BIOCHEMISTRY-MOSCOW, 2000, 65 (03) : 355 - 360
  • [34] Purification and properties of exopolyphosphatase isolated from Saccharomyces cerevisiae vacuoles
    Andreeva, NA
    Kulakovskaya, TV
    Kulaev, IS
    FEBS LETTERS, 1998, 429 (02) : 194 - 196
  • [35] Channel-forming activities of peroxisomal membrane proteins from the yeast Saccharomyces cerevisiae
    Grunau, Silke
    Mindthoff, Sabrina
    Rottensteiner, Hanspeter
    Sormunen, Raija T.
    Hiltunen, J. Kalervo
    Erdmann, Ralf
    Antonenkov, Vasily D.
    FEBS JOURNAL, 2009, 276 (06) : 1698 - 1708
  • [36] Function and regulation in MAPK signaling pathways:: Lessons learned from the yeast Saccharomyces cerevisiae
    Chen, Raymond E.
    Thorner, Jeremy
    BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2007, 1773 (08): : 1311 - 1340
  • [37] A simple method for culture and stable maintenance of giant spheroplasts from the yeast Saccharomyces cerevisiae
    Fukudomi, Taihei
    Kotani, Tomohiro
    Miyakawa, Isamu
    JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY, 2011, 57 (04) : 177 - 182
  • [38] Peptide extract from spent yeast improves resistance of Saccharomyces cerevisiae to oxidative stress
    Lopes, Ana
    Azevedo-Silva, Joao
    Carsanba, Erdem
    Pintado, Manuela
    Oliveira, Ana Sofia
    Ferreira, Carlos
    Pereira, Joana Odila
    Carvalho, Ana P.
    Oliveira, Carla
    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2023, 107 (11) : 3405 - 3417
  • [39] A dye-affinity cryogel membrane for malate dehydrogenase purification from Saccharomyces cerevisiae
    Hamade, Kamar
    Gokturk, Ilgim
    Bereli, Nilay
    Turkmen, Deniz
    Elkak, Assem
    Denizli, Adil
    JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 2020, 31 (01) : 38 - 52
  • [40] PURIFICATION AND FUNCTIONAL-CHARACTERIZATION OF THE PYRUVATE (MONOCARBOXYLATE) CARRIER FROM BAKERS-YEAST MITOCHONDRIA (SACCHAROMYCES-CEREVISIAE)
    NALECZ, MJ
    NALECZ, KA
    AZZI, A
    BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1079 (01) : 87 - 95