Difference between revisions of "Part:BBa K1216005"
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<partinfo>BBa_K1216001 short</partinfo> | <partinfo>BBa_K1216001 short</partinfo> | ||
| − | The alkaline phosphatase is a periplasmic homodimeric hydrolase. [[File:Alkaline_Phosphatase_3D.jpg|thumb| 3D representation of the alkaline phosphatase from [http://www.rcsb.org/pdb/explore/explore.do?structureId=1ANJ RCSB] ]] | + | The alkaline phosphatase is a periplasmic homodimeric hydrolase. The poly-HIS tag can be used for protein purification (IMAC)[[Part:BBa_K1216001#References|<sup>[1]</sup>]]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [[Part:BBa_K1216001#References|<sup>[2]</sup>]]. [[File:Alkaline_Phosphatase_3D.jpg|thumb| 3D representation of the alkaline phosphatase from [http://www.rcsb.org/pdb/explore/explore.do?structureId=1ANJ RCSB] ]] |
A form of this protein without TEV and poly-HIS tags can be found [[Part:BBa_K1216001| here]]. | A form of this protein without TEV and poly-HIS tags can be found [[Part:BBa_K1216001| here]]. | ||
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<!-- Add more about the biology of this part here--> | <!-- Add more about the biology of this part here--> | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | Alkaline phosphatases are used as reporter enzymes in different assays such as Western Blotting and in situ hybridization[[Part:BBa_K1216001#References|<sup>[ | + | Alkaline phosphatases are used as reporter enzymes in different assays such as Western Blotting and in situ hybridization[[Part:BBa_K1216001#References|<sup>[3]</sup>]]. Testing human blood for Alkaline Phosphatase levels is a routine test that can reveal different conditions[[Part:BBa_K1216001#References|<sup>[4]</sup>]]. |
| − | Alkaline phosphatases cleave phosphate groups from organic compounds by hydrolysis while retaining stereochemistry[[Part:BBa_K1216001#References|<sup>[ | + | Alkaline phosphatases cleave phosphate groups from organic compounds by hydrolysis while retaining stereochemistry[[Part:BBa_K1216001#References|<sup>[5]</sup>]]. |
A good explanation of the mechanism can be found [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates here].<br> | A good explanation of the mechanism can be found [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates here].<br> | ||
| − | Alkaline phosphatases, respectively their serum levels, are also related to several diseases e.g. metabolic myopathies and Paget Disease. [[Part:BBa_K1216001#References|<sup>[ | + | Alkaline phosphatases, respectively their serum levels, are also related to several diseases e.g. metabolic myopathies and Paget Disease. [[Part:BBa_K1216001#References|<sup>[6]</sup>]] |
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===References=== | ===References=== | ||
| + | # Loghran ST, "Purification of poly-histidine-tagged proteins.",Methods Mol Biol. 2011;681:311-35. doi: 10.1007/978-1-60761-913-0_17.[http://www.ncbi.nlm.nih.gov/pubmed/20978973] | ||
| + | # [http://homepage.univie.ac.at/nikos.pinotsis/tev_protease.html University of Vienna TEV Protease info] | ||
# Molecular Cell Biology, Fifth Edition, W.H. Freeman & Co., 2004. | # Molecular Cell Biology, Fifth Edition, W.H. Freeman & Co., 2004. | ||
# [http://www.nlm.nih.gov/medlineplus/ency/article/003470.htm Medline Plus] | # [http://www.nlm.nih.gov/medlineplus/ency/article/003470.htm Medline Plus] | ||
# [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates Section 10.3: Hydrolysis of phosphates] | # [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates Section 10.3: Hydrolysis of phosphates] | ||
# Adams & Victor's Principles Of Neurology, 7th edition, McGraw-Hill Professional, 2000. | # Adams & Victor's Principles Of Neurology, 7th edition, McGraw-Hill Professional, 2000. | ||
Revision as of 12:55, 6 September 2013
Alkaline Phosphatase (phoA) from Citrobacter
The alkaline phosphatase is a periplasmic homodimeric hydrolase. The poly-HIS tag can be used for protein purification (IMAC)[1]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [2].
A form of this protein without TEV and poly-HIS tags can be found here.
Usage and Biology
Alkaline phosphatases are used as reporter enzymes in different assays such as Western Blotting and in situ hybridization[3]. Testing human blood for Alkaline Phosphatase levels is a routine test that can reveal different conditions[4].
Alkaline phosphatases cleave phosphate groups from organic compounds by hydrolysis while retaining stereochemistry[5].
A good explanation of the mechanism can be found [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates here].
Alkaline phosphatases, respectively their serum levels, are also related to several diseases e.g. metabolic myopathies and Paget Disease. [6]
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 340
Illegal NgoMIV site found at 787 - 1000COMPATIBLE WITH RFC[1000]
References
- Loghran ST, "Purification of poly-histidine-tagged proteins.",Methods Mol Biol. 2011;681:311-35. doi: 10.1007/978-1-60761-913-0_17.[http://www.ncbi.nlm.nih.gov/pubmed/20978973]
- [http://homepage.univie.ac.at/nikos.pinotsis/tev_protease.html University of Vienna TEV Protease info]
- Molecular Cell Biology, Fifth Edition, W.H. Freeman & Co., 2004.
- [http://www.nlm.nih.gov/medlineplus/ency/article/003470.htm Medline Plus]
- [http://chemwiki.ucdavis.edu/Organic_Chemistry/Organic_Chemistry_With_a_Biological_Emphasis/Chapter_10%3A_Phosphoryl_transfer_reactions/Section_10.3%3A_Hydrolysis_of__phosphates Section 10.3: Hydrolysis of phosphates]
- Adams & Victor's Principles Of Neurology, 7th edition, McGraw-Hill Professional, 2000.