2any: Difference between revisions
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'''Expression, Crystallization and the Three-dimensional Structure of the Catalytic Domain of Human Plasma Kallikrein: Implications for Structure-Based Design of Protease Inhibitors''' | {{Structure | ||
|PDB= 2any |SIZE=350|CAPTION= <scene name='initialview01'>2any</scene>, resolution 1.40Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene> and <scene name='pdbligand=BAM:BENZAMIDINE'>BAM</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/Plasma_kallikrein Plasma kallikrein], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.34 3.4.21.34] | |||
|GENE= KLKB1, KLK3 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=9606 Homo sapiens]) | |||
}} | |||
'''Expression, Crystallization and the Three-dimensional Structure of the Catalytic Domain of Human Plasma Kallikrein: Implications for Structure-Based Design of Protease Inhibitors''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
2ANY is a [ | 2ANY is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2ANY OCA]. | ||
==Reference== | ==Reference== | ||
Expression, crystallization, and three-dimensional structure of the catalytic domain of human plasma kallikrein., Tang J, Yu CL, Williams SR, Springman E, Jeffery D, Sprengeler PA, Estevez A, Sampang J, Shrader W, Spencer J, Young W, McGrath M, Katz BA, J Biol Chem. 2005 Dec 9;280(49):41077-89. Epub 2005 Sep 30. PMID:[http:// | Expression, crystallization, and three-dimensional structure of the catalytic domain of human plasma kallikrein., Tang J, Yu CL, Williams SR, Springman E, Jeffery D, Sprengeler PA, Estevez A, Sampang J, Shrader W, Spencer J, Young W, McGrath M, Katz BA, J Biol Chem. 2005 Dec 9;280(49):41077-89. Epub 2005 Sep 30. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16199530 16199530] | ||
[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Plasma kallikrein]] | [[Category: Plasma kallikrein]] | ||
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[[Category: mutagenically deglycosyalted human plasma kallikrein protease domain; trypsin-like serine protease]] | [[Category: mutagenically deglycosyalted human plasma kallikrein protease domain; trypsin-like serine protease]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 15:51:51 2008'' | ||
Revision as of 16:51, 20 March 2008
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| , resolution 1.40Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Gene: | KLKB1, KLK3 (Homo sapiens) | ||||||
| Activity: | Plasma kallikrein, with EC number 3.4.21.34 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Expression, Crystallization and the Three-dimensional Structure of the Catalytic Domain of Human Plasma Kallikrein: Implications for Structure-Based Design of Protease Inhibitors
OverviewOverview
Plasma kallikrein is a serine protease that has many important functions, including modulation of blood pressure, complement activation, and mediation and maintenance of inflammatory responses. Although plasma kallikrein has been purified for 40 years, its structure has not been elucidated. In this report, we described two systems (Pichia pastoris and baculovirus/Sf9 cells) for expression of the protease domain of plasma kallikrein, along with the purification and high resolution crystal structures of the two recombinant forms. In the Pichia pastoris system, the protease domain was expressed as a heterogeneously glycosylated zymogen that was activated by limited trypsin digestion and treated with endoglycosidase H deglycosidase to reduce heterogeneity from the glycosylation. The resulting protein was chromatographically resolved into four components, one of which was crystallized. In the baculovirus/Sf9 system, homogeneous, crystallizable, and nonglycosylated protein was expressed after mutagenizing three asparagines (the glycosylation sites) to glutamates. When assayed against the peptide substrates, pefachrome-PK and oxidized insulin B chain, both forms of the protease domain were found to have catalytic activity similar to that of the full-length protein. Crystallization and x-ray crystal structure determination of both forms have yielded the first three-dimensional views of the catalytic domain of plasma kallikrein. The structures, determined at 1.85 A for the endoglycosidase H-deglycosylated protease domain produced from P. pastoris and at 1.40 A for the mutagenically deglycosylated form produced from Sf9 cells, show that the protease domain adopts a typical chymotrypsin-like serine protease conformation. The structural information provides insights into the biochemical and enzymatic properties of plasma kallikrein and paves the way for structure-based design of protease inhibitors that are selective either for or against plasma kallikrein.
DiseaseDisease
Known diseases associated with this structure: Fletcher factor deficiency OMIM:[229000], Prekallikrein deficiency OMIM:[229000]
About this StructureAbout this Structure
2ANY is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
ReferenceReference
Expression, crystallization, and three-dimensional structure of the catalytic domain of human plasma kallikrein., Tang J, Yu CL, Williams SR, Springman E, Jeffery D, Sprengeler PA, Estevez A, Sampang J, Shrader W, Spencer J, Young W, McGrath M, Katz BA, J Biol Chem. 2005 Dec 9;280(49):41077-89. Epub 2005 Sep 30. PMID:16199530
Page seeded by OCA on Thu Mar 20 15:51:51 2008
Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)
OCA- Pages with broken file links
- Homo sapiens
- Plasma kallikrein
- Single protein
- Estevez, A.
- Jeffery, D.
- Katz, B A.
- McGrath, M E.
- Sampang, J.
- Shrader, W.
- Spencer, J R.
- Sprengeler, P A.
- Springman, E.
- Tang, J.
- Williams, S R.
- Young, W B.
- Yu, C L.
- BAM
- PO4
- Mutagenically deglycosyalted human plasma kallikrein protease domain; trypsin-like serine protease