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Mass Spectrometry Analysis of Aquaporin-0
The lens fiber major intrinsic protein (otherwise known as aquaporin-0 (AQP0), MIP26 and MP26) has been examined by mass spectrometry (MS) in order to assay posttranslational modifications, particularly the speciation of acyl modifications to the side chains of lysine residues and the N-terminal amino group. The speciation of acyl modifications to the side chain of one specific, highly conserved lysine residue (K238) and the N-terminal amino group of human and bovine AQP0 revealed, in decreasing order of abun- dance, oleoyl, palmitoyl, stearoyl, eicosenoyl, dihomo-γ-linolenoyl, palmitoleoyl and eicosadienoyl modifications. In the case of human AQP0, an arachidonoyl modification was also found at the N-terminus. The relative abundances of these modifications mirror the fatty acid composition of lens phosphatidylethanolamine lipids. This lipid class would be ex- pected to be concentrated in the inner leaflet of the lens fiber membrane to which each of the potential AQP0 lipidation sites is proximal. Our data evidence a broad lipidation profile that is both species and site independent, suggesting a chemical-based ester aminolysis mechanism to explain such modifications.
Ministry of Higher Education and Scientific Research, Kurdistan
Fight for Sight UK
Data files are numbered (1-, 2- etc) according to the publication in which the data first appear. The next term is the species (Bov for bovine, Hum for human) and the lens section (Cortex or Nucleus).
Data are grouped according to the type of experiment: LC-MS, LC-MSMS, or LC-MSE (for trypsin digests) and Whole_Protein.
Data are presented in original (raw) format, and a more portable format (CDF or mzXML).
Those prefixed with '1' are from 10.1016/j.bbamem.2016.06.026
e.g. 1-Bov-Cortex_LC-MSMS_CDF.tbz2 is an archive containing data in CDF format from 10.1016/j.bbamem.2016.06.026 for the bovine lens cortex, analysed by LC-MSMS.
CDF files for Synapt G2-S data were generated using the Waters DataBridge program (as msconvert gave mass errors). DataBridge splits each raw data file into 4 CDF files. The LC-MS data are usually in the file suffixed with "01"' LC-MS/MS and LC-MSE data
are usually in the file suffixed with "02".
mzXML files generated using msconvert contain only the LC-MS portion of the data and may be truncated at
longer retention times (where there are no signals of interest).
LC-MSE data: contain within the filename the trap collision energy ramp (e.g. .....25_50_CE01.CDF for 25-50 eV).
LC-MSMS data: as LC-MSE data, plus the filename contains the mass of the parent ion (e.g. ...MSMS998....CE01.CDF for fragmentation of m/z 972)
LC-MS data: e.g. VSI_AQPO0C1_TRYP_DIG_DTT_SOD_AZID_C18_111214 was an outer cortex sample (C1) digested with trypsin, with a buffer containing DTT and sodium azide, run on a C18 column on 11th Dec 2014;
VSI_TRYPDIGEST_AQPO1_DTT_SODAZIDE_C18_220714 was an outer cortex sample (01)
digested with trypsin, with a buffer containing DTT and sodium azide, run on a C18 column on 22nd July 2014.
Whole (intact) Protein data: e.g. VSI_AQPO0C1_C4_SOD_AZ_PFOA_191214 was an outer cortex (01) sample, buffer containing sodium azide and PFOA, run on a C4 column on 19th Dec 2014.
LC-MS data: e.g. VSI_AQPO0N_TRYP_DIG_DTT_SODAZID_C18_NEW_121214 was an nuclear sample (N) digested with trypsin, with a buffer containing DTT and sodium azide, run on a C18 column on 12th Dec 2014.
LC-MSE data: contain within the filename the sample injection volume for the run (e.g. ...4UL....CE01.CDF for 4 microlitres).
LC-MSE data: contain within the filename the age and gender of the donor (e.g. ...M22....CE01.CDF for 22 year old male).
LC-MSE data: contain within the filename the trap collision energy ramp (e.g. .....30_50_CE01.CDF for 30-50 eV).
LC-MSMS data: as LC-MSE data, plus the filename contains the mass of the parent ion (e.g. ...MSMS972....CE01.CDF for fragmentation of m/z 972)
Output files for h-AQP0 are in the directory Human/HeliQuest.