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As explained in detail in the "Methods" section, this issue was addressed by using the protein identity group (PIG), which disambiguates different GIs that have an identical protein sequence.
Networks using the protein identity were generated using custom scripts and analyzed with CytoScape [ 50] and clusterMarker [ 51].
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25 Protein identities were determined using the Protein Global Server 1.0/2.0 software, and were further confirmed by Mascot (www.matrixscience.com) by using the MS/MS peak lists exported from MassLynx.
Using MALDI ToF MS, we determined the protein identity (Table 1) for 9 spots from the striatum and for 10 spots from the hippocampus, respectively.
However, in some cases two different algorithms, MS-Fit and Aldente, were used to provide support for the protein identity from the PMF in order to minimize false positive identifications.
We confirmed further the protein identity of selected spots (see "Results") using MALDI MS/MS analysis, using Bruker Ultraflex II.
The protein identity was confirmed by analysis using MALDI-TOF/TOF.
To confirm the protein identity, sequences were subjected to a profile search using Pfam [ 48].
Active AnMan5A, AnMan5B and AnMan5C were expressed in P. pastoris and purified from the culture supernatant using metal ion chromatography, and the protein identities were confirmed by mass spectrometry.
The protein identities reported were ranked high in at least two of the three algorithms used.
The obtained list of different m/ z ratios was used to verify protein identity using MS-Fit (ProteinProspector, UCSF; http://prospector.ucsf.edu/prospector/mshome.htm).htm
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