Proteomics

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Proteomic dissection of CFAP44 deficient axonemes


ABSTRACT: Leishmania are protists (class: Kinetoplastida) with a single multifunctional flagellum, which forms a canonical motile 9+2 microtubule axoneme in the promastigote forms. Deletion of gene CFAP44 (LmxM.14.1430) caused a reduction in promastigote motility (Beneke et al., PLoS Pathogens, 2019; accepted). To study the changes in protein composition in CFAP44 deficient axonemes, flagellar skeletons were isolated from CFAP44 knockout mutants and the parental cell line L. mex Cas9 T7 (Beneke et al., R Soc Open Sci. 2017; 4(5):170095) using a modified version of the method by Robinson et al., (Methods Enzymol. 1991; 196:285-99). Liquid chromatography tandem mass spectrometry and a label-free quantitation method (SINQ; Trudgian et al., 2011, Proteomics 10.1002) were used to identify proteins enriched in each fraction. This PRIDE upload contains .RAW and .XML files, as well as the SINQ quantification output file “SINQ_raw_data”. XML files are named SUB9810; MSS11680. .RAW files are structured as follows: “CFAP44 mutants: Qex01_SVH_180422_TomBeneke_B29_F_001”; “Cas9 T7 parentals: Qex01_SVH_180422_TomBeneke_Cas9_F_005”.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Leishmania Mexicana Mhom/gt/2001/u1103

TISSUE(S): Promastigote

SUBMITTER: Tom Beneke  

LAB HEAD: Dr Eva Gluenz

PROVIDER: PXD014077 | Pride | 2020-01-20

REPOSITORIES: Pride

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Publications

LAX28 is required for the stable assembly of the inner dynein arm f complex, and the tether and tether head complex in <i>Leishmania</i> flagella.

Beneke Tom T   Banecki Katherine K   Fochler Sophia S   Gluenz Eva E  

Journal of cell science 20200123 2


Motile eukaryotic flagella beat through coordinated activity of dynein motor proteins; however, the mechanisms of dynein coordination and regulation are incompletely understood. The inner dynein arm (IDA) f complex (also known as the I1 complex), and the tether and tether head (T/TH) complex are thought to be key regulators of dynein action but, unlike the IDA f complex, T/TH proteins remain poorly characterised. Here, we characterised T/TH-associated proteins in the protist <i>Leishmania mexica  ...[more]

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