Proteomic Analysis of Human Tears:  Defensin Expression after Ocular Surface Surgery

• Published in: Journal of proteome research Vol. 3; no. 3; pp. 410 - 416
• Main Authors: Zhou, L, Huang, L. Q, Beuerman, R. W, Grigg, M. E, Li, S. F. Y, Chew, F. T, Ang, L, Stern, M. E, Tan, D
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.05.2004
• Subjects: alpha-Defensins - analysis; alpha-Defensins - metabolism; Amino Acid Sequence; Chromatography, Liquid; Databases, Protein; Eye Neoplasms - metabolism; Eye Neoplasms - surgery; Eye Proteins - analysis; Eye Proteins - metabolism; Humans; Lasers; Mass Spectrometry; Molecular Sequence Data; Tears - metabolism
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr034065n

Human tear protein profiles were monitored by surface enhanced laser desorption/ionization-time-of-flight mass spectrometry ProteinChip technology (SELDI-TOF ProteinChip) and liquid chromatography−mass spectrometry (LC−MS). Tears were collected from 21 patients scheduled for surgery to remove an ocular surface neoplasm prior to surgery (day 0) and on days 1, 3, and 30 postoperatively. Using this proteomic approach, we verified that three human α-defensins (HNP-1, HNP-2, and HNP-3) were significantly up-regulated in their expression after surgery and that their levels decreased to approximately normal by day 30 by which time healing was complete. Further confirmation of the identity of the α-defensins in human tears was made by LC purification, trypsin digestion, and ESI−MS/MS analysis of their tryptic digests. The concentrations of HNP-1 and HNP-2 were determined and shown to be markedly increased after ocular surface surgery. The results of the study suggest that human α-defensins HNP-1, HNP-2, and HNP-3 are up-regulated after surgery, and may in addition to their antimicrobial properties have an important role in wound healing. Keywords: human tear proteins • eye • defensins • SELDI-TOF • ProteinChip • proteomics