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Evidence for existence of thirty hypothetical proteins in rat brain.

Shin JH, Yang JW, Juranville JF, Fountoulakis M, Lubec G - Proteome Sci (2004)

Bottom Line: RESULTS: We constructed a two-dimensional protein map and searched for expression of hypothetical proteins in rat brain.A number of hypothetical gene products were detected (30 of 190, 15.8%) and are considered brain proteins.CONCLUSIONS: A major finding of this study is the demonstration of the existence of putative proteins that were so far only deduced from their nucleic acid structure by a protein chemical method independent of antibody availability and specificity and unambiguously identifying proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, University of Vienna, Vienna, Austria. gert.lubec@akh-wien.ac.at

ABSTRACT
BACKGROUND: The rapid completion of genome sequences has created an infrastructure of biological information and provided essential information to link genes to gene products, proteins, the building blocks for cellular functions. In addition, genome/cDNA sequences make it possible to predict proteins for which there is no experimental evidence. Clues for function of hypothetical proteins are provided by sequence similarity with proteins of known function in model organisms. RESULTS: We constructed a two-dimensional protein map and searched for expression of hypothetical proteins in rat brain. Two-dimensional electrophoresis (2-DE) with subsequent in-gel digestion of spots and matrix-assisted laser desorption/ionization (MALDI) spectrometric identification were applied. In total about 3700 spots were analysed, which resulted in the identification of about 1700 polypeptides, that were the products of 190 different genes. A number of hypothetical gene products were detected (30 of 190, 15.8%) and are considered brain proteins. CONCLUSIONS: A major finding of this study is the demonstration of the existence of putative proteins that were so far only deduced from their nucleic acid structure by a protein chemical method independent of antibody availability and specificity and unambiguously identifying proteins.

No MeSH data available.


Related in: MedlinePlus

2-DE gel image of rat brain proteins depicting identified 30 hypothetical proteins. Accession numbers are given. Brain proteins were extracted and separated on an immobilised pH 3–10 non-linear gradient strip followed by separation on a 9–16% gradient polyacrylamide gel. The gel was stained with Coomassie blue and spots were analysed by MALDI-MS.
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Figure 1: 2-DE gel image of rat brain proteins depicting identified 30 hypothetical proteins. Accession numbers are given. Brain proteins were extracted and separated on an immobilised pH 3–10 non-linear gradient strip followed by separation on a 9–16% gradient polyacrylamide gel. The gel was stained with Coomassie blue and spots were analysed by MALDI-MS.

Mentions: Rat brain proteins were solubilised in the IEF-compatible reagents urea, thiourea and CHAPS and analysed by 2-D gels. The 2-DE separation was performed on broad pH range IPG strips and protein spots were visualised following staining with colloidal Coomassie blue. A large series of proteins were successfully identified, including hypothetical proteins. A representative gel presenting hypothetical proteins is shown in Fig. 1.


Evidence for existence of thirty hypothetical proteins in rat brain.

Shin JH, Yang JW, Juranville JF, Fountoulakis M, Lubec G - Proteome Sci (2004)

2-DE gel image of rat brain proteins depicting identified 30 hypothetical proteins. Accession numbers are given. Brain proteins were extracted and separated on an immobilised pH 3–10 non-linear gradient strip followed by separation on a 9–16% gradient polyacrylamide gel. The gel was stained with Coomassie blue and spots were analysed by MALDI-MS.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC373456&req=5

Figure 1: 2-DE gel image of rat brain proteins depicting identified 30 hypothetical proteins. Accession numbers are given. Brain proteins were extracted and separated on an immobilised pH 3–10 non-linear gradient strip followed by separation on a 9–16% gradient polyacrylamide gel. The gel was stained with Coomassie blue and spots were analysed by MALDI-MS.
Mentions: Rat brain proteins were solubilised in the IEF-compatible reagents urea, thiourea and CHAPS and analysed by 2-D gels. The 2-DE separation was performed on broad pH range IPG strips and protein spots were visualised following staining with colloidal Coomassie blue. A large series of proteins were successfully identified, including hypothetical proteins. A representative gel presenting hypothetical proteins is shown in Fig. 1.

Bottom Line: RESULTS: We constructed a two-dimensional protein map and searched for expression of hypothetical proteins in rat brain.A number of hypothetical gene products were detected (30 of 190, 15.8%) and are considered brain proteins.CONCLUSIONS: A major finding of this study is the demonstration of the existence of putative proteins that were so far only deduced from their nucleic acid structure by a protein chemical method independent of antibody availability and specificity and unambiguously identifying proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, University of Vienna, Vienna, Austria. gert.lubec@akh-wien.ac.at

ABSTRACT
BACKGROUND: The rapid completion of genome sequences has created an infrastructure of biological information and provided essential information to link genes to gene products, proteins, the building blocks for cellular functions. In addition, genome/cDNA sequences make it possible to predict proteins for which there is no experimental evidence. Clues for function of hypothetical proteins are provided by sequence similarity with proteins of known function in model organisms. RESULTS: We constructed a two-dimensional protein map and searched for expression of hypothetical proteins in rat brain. Two-dimensional electrophoresis (2-DE) with subsequent in-gel digestion of spots and matrix-assisted laser desorption/ionization (MALDI) spectrometric identification were applied. In total about 3700 spots were analysed, which resulted in the identification of about 1700 polypeptides, that were the products of 190 different genes. A number of hypothetical gene products were detected (30 of 190, 15.8%) and are considered brain proteins. CONCLUSIONS: A major finding of this study is the demonstration of the existence of putative proteins that were so far only deduced from their nucleic acid structure by a protein chemical method independent of antibody availability and specificity and unambiguously identifying proteins.

No MeSH data available.


Related in: MedlinePlus