Proteomics

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Extraction of Entamoeba histolytica membrane proteins using three extraction methods

ABSTRACT: Entamoeba histolytica membrane proteins are important players in the parasite’s pathogenicity. However, most of the proteins have not been identified. This study reports the membrane proteins extracted using three extractions methods: two commercial kits (ProteoExtract® from Calbiochem and ProteoPrep® from Sigma), and a conventional laboratory method. The resulting membrane fractions (MF) and cytosolic fractions (CF)were analysed using LC-ESI-MS/MS. The proteins identified in at least two out of three biological replicates revealed a total of 490, 492, and 587 MF proteins extracted using the ProteoExtract® kit, ProteoPrep® kit and conventional method, respectively. Meanwhile, 487, 611 and 343 proteins were identified in the CF extracted using the ProteoExtract® kit, ProteoPrep® kit and conventional method, respectively. Analysis of the identified MF and CF proteins extracted by the respective extraction kits suggests that the ProteoPrep® extraction kit was the most selective in separating MF and CF among the three extraction methods.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Entamoeba Histolytica Hm-1:imss

SUBMITTER: JORIM UJANG  

LAB HEAD: Nurulhasanah binti Othman

PROVIDER: PXD010171 | Pride | 2018-10-16

REPOSITORIES: Pride

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Source:
Action DRS
ConventionalCF1mgf.mgf Mgf
ConventionalCF1msf.msf Msf
ConventionalCF1raw.raw Raw
ConventionalCF2mgf.mgf Mgf
ConventionalCF2msf.msf Msf
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Analysis of Entamoeba histolytica Membrane Proteome Using Three Extraction Methods.

Ujang Jorim J   Sani Asmahani Azira Abdu AAA   Lim Boon Huat BH   Noordin Rahmah R   Othman Nurulhasanah N  

Proteomics 20181025 23

Entamoeba histolytica membrane proteins are important players toward the pathogenesis of amoebiasis, but the roles of most of the proteins are not fully understood. Since efficient protein extraction method is crucial for a successful MS analysis, three extractions methods are evaluated for the use in studying the membrane proteome of E. histolytica: Two commercial kits (ProteoExtract from Calbiochem and ProteoPrep from Sigma), and a conventional laboratory method. The results show that ProteoEx  ...[more]

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