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Resolution ofFusarium sporotrichioides proteins by two-dimensional polyacrylamide gel electrophoresis and identification by sequence homology comparison in protein data base (1995)

Chow, Lu-Ping ; Fukaya, Nobuhito ; Miyatake, Norifumi ; [et al.]

Springer

Journal of biomedical science 2 (1995), S. 343-352

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ISSN: 1423-0127

Keywords: Two-dimensional gel electrophoresis ; Protein separation ; Fusarium sporotrichioides ; Imperfect fungus

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Abstract Proteins fromFusarium sporotrichioides M-1-1, a T2-toxin-producing strain, were separated by two-dimensional polyacrylamide gel electrophoresis. One thousand two hundred and forty-four protein spots were resolved and 103 protein spots were subjected to N-terminal sequencing. Fifty-eight protein spots were sequenced and 48 proteins were observed to have blocked N termini. Forty out of 58 sequenced proteins were identified by homology search against the PIR protein sequence data base and protein superfamily data base, while the residual 18 sequences were not identified. Twenty-seven of the N-terminal-blocked proteins were subjected to mild anhydrous hydrazine vapor deblocking. Twenty-four spots were not deblocked indicating the presence of acyl groups at the N termini, while 3 proteins were deblocked showing the blocked group to be pyrroglutamyl carboxylic acid residues. The results can provide a more global view of cellular genetic expression than any other technique. The created data may offer a unique opportunity to link information with DNA sequence data.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF02255221

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Separation and characterization of Arabidopsis thaliana proteins by two-dimensional gel electrophoresis (1995)

Kamo, Masaharu ; Kawakami, Takao ; Miyatake, Norifumi ; [et al.]

Weinheim : Wiley-Blackwell

Electrophoresis 16 (1995), S. 423-430

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ISSN: 0173-0835

Keywords: Two-dimensional polyacrylamide gel electrophoresis ; Arabidopsis thaliana ; Gel image analysis ; N-Terminal sequences ; Deblocking by hydrazine vapor ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Chemistry and Pharmacology

Notes: Arabidopsis (Arabidopsis thaliana) proteins were isolated from five tissues (leaf, stem, root, seed and callus), and separated by two-dimensional gel electrophoresis (2-DE). 2-DE was carried out by immobilized pH gradient (IPG) in the first dimension, and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. With the aid of comigrated five-marker proteins, the patterns of 2-DE gels for each tissue were graphically combined by a computer into a single synthetic image for the integrated Arabidopsis protein spots. The protein spot images, altogether 4763, were characterized by both molecular mass and isoelectric point. Partial amino(N)-terminal sequences of 101 protein spots were analyzed by Edman degradation. Fifty seven proteins were partially sequenced and 46 proteins appeared to have blocked N-termini. Deblocking by hydrazine vapor was carried out on 14 proteins and two of them were found to be pyroglutamyl-blocked N-termini. Forty seven new proteins were found by the present investigation.

Additional Material: 2 Ill.