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. 2014 Nov 13;9(11):e111645.
doi: 10.1371/journal.pone.0111645. eCollection 2014.

Quantitative proteomic analysis of serum from pregnant women carrying a fetus with conotruncal heart defect using isobaric tags for relative and absolute quantitation (iTRAQ) labeling

Ying Zhang  1 , Yuan Kang  1 , Qiongjie Zhou  1 , Jizi Zhou  1 , Huijun Wang  2 , Hong Jin  3 , Xiaohui Liu  4 , Duan Ma  5 , Xiaotian Li  6
Affiliations

Quantitative proteomic analysis of serum from pregnant women carrying a fetus with conotruncal heart defect using isobaric tags for relative and absolute quantitation (iTRAQ) labeling

Ying Zhang et al. PLoS One. .

Abstract

Objective: To identify differentially expressed proteins from serum of pregnant women carrying a conotruncal heart defects (CTD) fetus, using proteomic analysis.

Methods: The study was conducted using a nested case-control design. The 5473 maternal serum samples were collected at 14-18 weeks of gestation. The serum from 9 pregnant women carrying a CTD fetus, 10 with another CHD (ACHD) fetus, and 11 with a normal fetus were selected from the above samples, and analyzed by using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry(2D LC-MS/MS). The differentially expressed proteins identified by iTRAQ were further validated with Western blot.

Results: A total of 105 unique proteins present in the three groups were identified, and relative expression data were obtained for 92 of them with high confidence by employing the iTRAQ-based experiments. The downregulation of gelsolin in maternal serum of fetus with CTD was further verified by Western blot.

Conclusions: The identification of differentially expressed protein gelsolin in the serum of the pregnant women carrying a CTD fetus by using proteomic technology may be able to serve as a foundation to further explore the biomarker for detection of CTD fetus from the maternal serum.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Flow diagram describing the selection of the analytical data set used for iTRAQ analysis.
Figure 2
Figure 2. Identification of the protein gelsolin as being differentially expressed in maternal sera of pregnant women carrying a CTD fetus.
(A) Identification of the protein gelsolin in maternal serum by iTRAQ and 2D LC-MS/MS. (i) The peptide quantitation information of gelsolin was derived from the intensities of three iTRAQ reporter ions. Reporter ions 118,119 were used to label sera from pregnant women carrying CTD and ACHD fetuses, respectively. Reporter ion 121 was used to label normal controls. (ii)The representative MS/MS spectrum of a peptide from gelsolin. (B). The Western blot validated the relative expression of GSN, A2M, CERU, ATRN, PZP in maternal serum samples(i), and confirmed relative decreased level of maternal serum gelsolin in CTD group compared with normal controls (ii). n = 9 in CTD group, n = 11 in normal control.*p = 0.008.
Figure 3
Figure 3. Western blot and IHC analyses confirmed the relative expression of GSN in fetal heart tissues with CTD.
(A) Western blot analysis examined the relative expression of GSN, and confirmed that the GSN protein was downregulated in heart tissues of CTD fetuses compared with normal controls. n = 4, respectively. *p = 0.004. (B) IHC demonstrated the dark brown immunostaining was weaker in the CTD heart tissues than the normal controls (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks). n = 4, respectively. Magnification was set at 400×.
See this image and copyright information in PMC

References

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