OBJECTIVES: Ankylosing spondylitis (AS) is difficult to diagnose in its early stage due to the lack of simple and specific diagnostic indicators. This study was performed to screen candidate AS-associated proteins from peripheral blood mononuclear cells (PBMCs) of AS patients by combining a two-dimensional electrophoresis (2-DE) technique with mass spectrometry (MS) analysis. METHODS: Twelve subjects consisting of 6 AS patients and 6 healthy volunteers (HVs) were enrolled in the 2-DE experiments. The protein expression patterns of PBMCs from different groups were analysed by PDQuest software, and the protein spots over/under-expressed by more than 2-fold between the two groups were identified by MS analysis. Western blot analyses were used to verify the differentially expressed proteins in 32 AS patients and 32 HVs. RESULTS: Six proteins including pyruvate kinase (PK), profilin 1 (PFN1), talin 1 (TLN1), Chain A of cyclophilin A (CyPA), unknown protein (gi|16306948) and integrin-linked kinase (ILK) were identified from 10 over-expressed protein spots found by 2-DE in the AS group. Western blot experiments confirmed a higher expression of both TLN1 and ILK in AS group compared to the HV group (p<0.05). CONCLUSIONS: TLN1 and ILK expressed higher in PBMCs of AS patients compared to healthy controls, which were involved in the integrin signalling pathway. The two proteins are likely novel disease-associated proteins and potential disease markers of AS.
OBJECTIVES:Ankylosing spondylitis (AS) is difficult to diagnose in its early stage due to the lack of simple and specific diagnostic indicators. This study was performed to screen candidate AS-associated proteins from peripheral blood mononuclear cells (PBMCs) of AS patients by combining a two-dimensional electrophoresis (2-DE) technique with mass spectrometry (MS) analysis. METHODS: Twelve subjects consisting of 6 AS patients and 6 healthy volunteers (HVs) were enrolled in the 2-DE experiments. The protein expression patterns of PBMCs from different groups were analysed by PDQuest software, and the protein spots over/under-expressed by more than 2-fold between the two groups were identified by MS analysis. Western blot analyses were used to verify the differentially expressed proteins in 32 AS patients and 32 HVs. RESULTS: Six proteins including pyruvate kinase (PK), profilin 1 (PFN1), talin 1 (TLN1), Chain A of cyclophilin A (CyPA), unknown protein (gi|16306948) and integrin-linked kinase (ILK) were identified from 10 over-expressed protein spots found by 2-DE in the AS group. Western blot experiments confirmed a higher expression of both TLN1 and ILK in AS group compared to the HV group (p<0.05). CONCLUSIONS:TLN1 and ILK expressed higher in PBMCs of AS patients compared to healthy controls, which were involved in the integrin signalling pathway. The two proteins are likely novel disease-associated proteins and potential disease markers of AS.
Authors: Roman Fischer; David C Trudgian; Cynthia Wright; Gethin Thomas; Linda A Bradbury; Matthew A Brown; Paul Bowness; Benedikt M Kessler Journal: Mol Cell Proteomics Date: 2011-10-13 Impact factor: 5.911