Phd Thesis 33 - eCommons@USASK - University of Saskatchewan

(Figure 4.5 continued)
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McAPN1 ----------------------------------------------------
HaAPN6 ----------------------------------------------------
McAPN3 ----------------------------------------------------
HaAPN4 ----------------------------------------------------
McAPN4 SVSTEAPTTEATTTPEPSSAAIYMPTTMLLLWTALAMLLK------------
HaAPN2 TEGSGEESTTPDGSSTTAAPESGESEQDESSSATIFLPTTVLLSWTVLAMLL
McAPN6 FAHIIL----------------------------------------------
HaAPN3 SLGAILHLII------------------------------------------
McAPN7 ASSTTAAPITEPSTPEPTTTPAPDSANLASLSIVTLIVTLVINMA-------
HaAPN1 MIVTLVVNMA------------------------------------------
McAPN8 ----------------------------------------------------
HaAPN5 ----------------------------------------------------
Figure 4.5 Alignment of Mamestra configurata peritrophic matrix aminopeptidases (APNs) with
those from H. armigera. Location of the signal peptides (bold, italics and underlined), zinc
binding site “HEXXH” (green lettering in green box), putative glycosyl phosphatidylinositol
modification sites (orange lettering, underlined), asparagines that were predicted to be N-
glycosylated (blue lettering), or O-glycosylated amino acids (white lettering) within the serinethreonine-proline
rich region (green background) and amino acids present in the majority (gray
background are shown. Accession numbers of the HaAPNs are: HaAPN1 (ACC68683), HaAPN2
(AAP37951), HaAPN3 (AAP37953), HaAPN4 (AAP37950), HaAPN5 (ABN64103), and
HaAPN6 (ACA35025).
Mascot analysis identified one to seven peptide hits linked to five putative
carboxypeptidases (with a peptidase M14 carboxypeptidase domain) (Marchler-Bauer et al.,
2009) (Table 4.2, Appendix A). These include three A Type carboxypeptidases, CPA, (F 255 , V 255
and S 255 ) [McCPA2, 5 and 6, respectively] and two B Type carboxypeptidases, CPB, (R 255 and
E 255 ) [McCPB1 and CPB4, respectively] (Figure 4.6). McCPB1, McCPA2 and McCPB4 were
identified previously (Hegedus et al., 2003; Erlandson et al., 2010). The two new
carboxypeptidases, McCPA5 and CPA6, had partial sequences with a 665 bp cDNA encoding a
200 amino acid protein and a 682 bp cDNA encoding a 226 amino acid protein, respectively. All
M.configurata carboxypeptidases that were found to be PM associated had a signal peptide, an
arginine as a putative trypsin cleavage site near their amino termini, a zinc binding motif (H 69 ,
E 72 and H 196 , numbering is according to the Bos taurus carboxypeptidases) (except McCPA5)
and one or two N-glycosylated asparagines (except McCPA2). In addition, a pair of conserved
cysteines (C 138 and C 161 ) was found in McCPB1, McCPB4 and McCPA6; however, McCPA2
and McCPA5 lacked at least one of these. These features are shown in an alignment of McCPs
with the model CPA (P00730) and CPB (P00732) from B. taurus (Figure 4.6).
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