Spatial differences in an integral membrane proteome detected in laser capture microdissected samples.
The combination of laser capture microdissection and mass spectrometry represents a powerful technology for studying spatially resolved proteomes. Moreover, the compositions of integral membrane proteomes have rarely been studied in a spatially resolved manner. In this study, ocular lens tissue was carefully dissected by laser capture microdissection and conditions for membrane protein enrichment, trypsin digestion, and mass spectrometry analysis were optimized. Proteomic analysis allowed the identification of 170 proteins, 136 of which were identified with more than one peptide match. Spatial differences in protein expression were observed between cortical and nuclear samples. In addition, the spatial distribution of post-translational modifications to lens membrane proteins, such as the lens major intrinsic protein AQP0, were investigated and regional differences were measured for AQP0 C-terminal phosphorylation and truncation.
Wang
Zhen
Z
Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Han
Jun
J
Schey
Kevin L
KL
D000818:Animals; D020346:Aquaporins / Q000378:metabolism; D002417:Cattle; D002853:Chromatography, Liquid; D017630:Connexins / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism*; D007834:Lasers; D007908:Lens, Crystalline / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D042282:Microdissection; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D000099264:Connexin 50
D020346:Aquaporins; D017630:Connexins; D005136:Eye Proteins; D008565:Membrane Proteins; D020543:Proteome; D000099264:Connexin 50; C030328:aquaporin 0; C082917:GJA3 protein, human