1

123

EdmanNMALDI/TOF/MSESI/MS/MSPSD/MALDI/TOF/MS32

11 SmithiesPoulik(1956)OFarrell1975()(SDSPAGE)

2 SDS-PAGE20cm10010 00030cm x 40 cm(20 cm x 20 cm)3000.

FLA-5000-image: Sypro Ruby stained 2 D-gel, brain proteins

21():

2(change coupled devices,CCD)

Image Scanner 14-bit3.7OD 1D2D

1000dpi(dot per inch)dpi300dpi8bits20cm x 20cm(Protean IIBio-Rad)tif 6Mb 300dpi300dpi256256

3 melanie II&3&4PDQuest 6.1ImageMaster 2D Elite 3.10Phoretix 2DNonlinear Dynamics Ltd.Progenesis Decodon GmbHDelta2D: spot detection background subtraction (normalization) (spot matching)

31 (m/z) 1886Goldstein194280fmol(10-15)amol(10-18).

2002

191719401959

2

1.2.3.1.2.3.4. 1. 2. 3.4.

3(Matrix-assisted laser desorption/ionization, MALDI)Elaectrospray Ionization-ESI

MALDI ()(337nm)

2) ESI N2

4(triple-quadrupole analyzer)(ion trap analyzer)(quadruple-time of light analyzer)

(time of flightTOF) (mz)

MALDI-TOF-MS2(RETOF-MS)

5MALDI-TOF/MS;ESI-Q-TOF/MS;MALDI-TOF-TOF/MS;ESI-MS/MS;HPLC-ESI-MS/MS.

MALDI-TOF/MS

HPLC-ESI-MS/MS

6A: (Y axis)B: (m/z, X axis)C: (base peak)D: E: m/z.F: (centroidal peak) G: (gauss peak)

MALDI-TOF/MS

LC-ESI-MS/MS

4(attribute parameter)(peptide mass fingerprintingPMF)MS/MS

1PMF1PMF1993()PVDFPMF

2

3

4Fig of Ap73a

Result of Ap73a

1,,() ,,:; ; ; ;

2,,,

1

,,,,,,Donohoe 151 ,;

,,,Biplab ,(mammalian target of rapamycin ,mTOR) ,;mTOR ,,, mTOR ,

2,,,,,,Graves ,1 (ALDH1) 2 (QR2),, QR2 ,

3,,,,,, ,Charlwood ,,

3

(2006) MALDI-TOFADP- (AGPase)UDP- A

(2002)40kD6.3 P40P40 Campo (2004)9 elF-5A

4

1

2

3

4

IgA / /E-mail:zhangyl@stu.edu.cn

IgIgA

IgAIgAIgSFIgAIgAIgA

3.1 IgA

3.2 1-DE/2-DE and 1-D/2-D Immunoblotting

Fig.1, 77 kDa, 75 kDa (Ap75) 64 kDa 3Ap75IgA-like) IgAAp75

3.3 Ap75 1 MALDI-TOF/MS

2ESI-MS/MS

Table 1: Matched amino acid sequences of Ap75 identified as L. vannamei hemocyanin by ESI-MS/MS analysis.

:Fig.24Table.1 MALDI-TOF/MS ESI-MS/MS Ap7575kDaIgA75kDa IgA

3.4 IgA175kDaConserved domain NCBIBLAST IgA75kDaC252Ig-like domain 75kDaIg-like domainIgAIg-like domain

2Ig-likeImmunoblotting

Fig5775kDaIgAIg-like domainIgSFIgIgSFIgSF

3.5 41-DE1SDS-PAGE

2MALDI-TOF

Fig.9 Table.2 4

3.6 6

Fig.10 Table.34

3.7 1 K12LPS

2 k12

3 OmpCOmpXK12OmpCOmpXOmpXOmpCOmpX

IgA1IgAIgA75kDa275kDaIgIg-like domain3IgIgSF4 6 5OmpX

1. Zhang, Y.L., Wang, S.Y., Xu, A.L., Chen, J., Lin, B.K., Peng, X.X. Affinity proteomic approach for identification of an IgA-like protein in Litopenaeus vannamei and study on its agglutination characterization.J.Proteome Res. 2006, 5:815-821 (IF, 6.92)2., , , . . , 2006, 13(6):1006-10113., . . 4. , . .