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Published on: October 19, 2014
Proteomic analysis of lymph
Lee V Leak1, Lance A Liotta, Henry Krutzsch
1Clinical Proteomics Program of Therapeutic Proteins, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA.
Insights
This study reveals that ovine lymph contains unique proteins, differing from blood plasma. Proteomic analysis identified specific lymph proteins, highlighting lymph
Area of Science:
- Proteomics
- Animal Science
- Biochemistry
Background:
- Lymph is traditionally viewed as an ultrafiltrate of blood plasma.
- Understanding the lymph proteome is crucial for comprehending its physiological roles.
- Previous analyses have not fully characterized the protein composition of normal ovine lymph.
Purpose of the Study:
- To conduct the first proteomic analysis of normal ovine lymph.
- To identify and differentiate proteins present in ovine lymph compared to blood plasma.
- To characterize the lymph proteome using advanced mass spectrometry techniques.
Main Methods:
- Utilized surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) with weak cation exchange chips.
- Employed two-dimensional gel electrophoresis (2-D PAGE) for protein separation.
- Conducted tandem mass spectrometry (MS/MS) for protein identification against a comprehensive database.
Main Results:
- SELDI-TOF-MS identified three protein peaks exclusive to lymph and five exclusive to plasma, with eight common peaks.
- 2-D PAGE confirmed higher ion presence in plasma than lymph.
- MS/MS identified 18 proteins in ovine lymph, including albumin, fibrinogen, immunoglobulin G (IgG), and serotransferrin. Glial fibrillary astrocyte acidic protein and neutrophil cytosol factor-1 were differentially expressed.
Conclusions:
- Ovine lymph possesses a distinct proteome, differing quantitatively and qualitatively from blood plasma.
- Proteomic technologies enable the detection of differentially expressed proteins in lymph.
- This study establishes lymph as more than a simple ultrafiltrate of plasma.
Abstract:
This report provides the first proteomic analysis of normal ovine lymph. By establishing the fact that lymph is more than an ultrafiltrate of blood plasma, it documents that the lymph proteome contains an array of proteins that differentiates it from plasma. The protein chip technology, surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS), two-dimensional gel electrophoresis (2-D PAGE) and MS, were employed to examine the protein expression profiles of ovine lymph. Using a weak cation exchange chip surface to assay lymph and plasma samples by SELDI-TOF-MS showed that the analysis of peak maps from lymph contained three protein peaks that were found only in lymph, while analysis of peak maps from plasma samples showed that five protein peaks were found only in plasma. Lymph and plasma samples showed eight peaks that were common to both. There were also more ions present in plasma than in lymph, which is consistent with the 2-D PAGE analysis. MS analysis of a large number of protein spots from 2-D PAGE gels of lymph produced MS/MS sequences for 18 proteins that were identified by searching against a comprehensive protein sequence database. As in plasma, large protein spots of albumin dominated the protein pattern in lymph. Other major proteins identified in 2-D PAGE gels of lymph included, fibrinogen alpha- and beta-chains, immunoglobulin G (IgG) heavy chain, serotransferrin precursor, lactoferrin, and apolipoprotein A-1. Two proteins that were identified and were differentially expressed in lymph were glial fibrillary astrocyte acidic protein and neutrophil cytosol factor-1. By bringing the technologies of proteomics to bear on the analysis of lymph, it is possible to detect proteins in lymph that are quantitatively and qualitatively differentially expressed from those of plasma.
