5vbu: Difference between revisions

Line 1:

~~'''Unreleased structure'''~~

~~The entry~~ 5vbu is ~~ON HOLD until Paper~~ Publication

==Crystal Structure of Human Cytochrome P450 21A2 Hydroxyprogesterone Complex==

== Structural highlights ==

<table><tr><td colspan='2'>[[5vbu]] is a 3 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5VBU OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5VBU FirstGlance]. <br>

</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=3QZ:(9BETA)-17-HYDROXYPREGN-4-ENE-3,20-DIONE'>3QZ</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>

<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5vbu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5vbu OCA], [http://pdbe.org/5vbu PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5vbu RCSB], [http://www.ebi.ac.uk/pdbsum/5vbu PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5vbu ProSAT]</span></td></tr>

</table>

== Publication Abstract from PubMed ==

Cytochrome P450 (P450, CYP) 21A2 is the major steroid 21-hydroxylase, converting progesterone to 11-deoxycorticosterone and 17alpha-hydroxyprogesterone (17alpha-OH-progesterone) to 11-deoxycortisol. More than 100 CYP21A2 variants give rise to congenital adrenal hyperplasia (CAH). We previously reported a structure of WT human P450 21A2 with bound progesterone and now present a structure bound to the other substrate (17alpha-OH-progesterone). We found that the 17alpha-OH-progesterone- and progesterone-bound complex structures are highly similar, with only some minor differences in surface loop regions. Twelve P450 21A2 variants associated with either salt-wasting or nonclassical forms of CAH were expressed, purified, and analyzed. The catalytic activities of these 12 variants ranged from 0.00009% to 30% of WT P450 21A2 and the extent of heme incorporation from 10% to 95% of the WT. Substrate dissociation constants (Ks) for four variants were 37-13,000-fold higher than for WT P450 21A2. Cytochrome b5, which augments several P450 activities, inhibited P450 21A2 activity. Similar to the WT enzyme, high noncompetitive intermolecular kinetic deuterium isotope effects (>/= 5.5) were observed for all six P450 21A2 variants examined for 21-hydroxylation of 21-d3-progesterone, indicating that C-H bond breaking is a rate-limiting step over a 104-fold range of catalytic efficiency. Using UV-visible and CD spectroscopy, we found that P450 21A2 thermal stability assessed in bacterial cells and with purified enzymes differed among salt-wasting- and nonclassical-associated variants, but these differences did not correlate with catalytic activity. Our in-depth investigation of CAH-associated P450 21A2 variants reveals critical insight into the effects of disease-causing mutations on this important enzyme.

~~Authors:~~ Pallan, P.S., ~~Egli, M~~.

Functional analysis of human cytochrome P450 21A2 variants involved in congenital adrenal hyperplasia.,Wang C, Pallan PS, Zhang W, Lei L, Yoshimoto FK, Waterman MR, Egli M, Guengerich FP J Biol Chem. 2017 Jun 30;292(26):10767-10778. doi: 10.1074/jbc.M117.792465. Epub , 2017 May 24. PMID:28539365<ref>PMID:28539365</ref>

~~Description: Crystal Structure~~ of ~~Human Cytochrome P450 21A2 Hydroxyprogesterone Complex~~

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>

~~[[Category~~: ~~Unreleased Structures]]~~

</div>

~~[[Category: Pallan, P.S]]~~

== References ==

__TOC__

</StructureSection>

[[Category: Egli, M]]

[[Category: Pallan, P S]]

[[Category: Addison's disease]]

[[Category: Adrenal steroidogenesis]]

[[Category: Congenital adrenal hyperplasia]]

[[Category: Hydroxyprogesterone]]

[[Category: Kinetic isotope effect]]

[[Category: Monooxygenase]]

[[Category: Oxidoreductase]]

[[Category: Steroid hydroxylation]]

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

Tools

@@ Line 1: / Line 1: @@
-'''Unreleased structure'''
-The entry 5vbu is ON HOLD  until Paper Publication
+==Crystal Structure of Human Cytochrome P450 21A2 Hydroxyprogesterone Complex==
+<StructureSection load='5vbu' size='340' side='right' caption='[[5vbu]], [[Resolution|resolution]] 3.31&Aring;' scene=''>
+== Structural highlights ==
+<table><tr><td colspan='2'>[[5vbu]] is a 3 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5VBU OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5VBU FirstGlance]. <br>
+</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=3QZ:(9BETA)-17-HYDROXYPREGN-4-ENE-3,20-DIONE'>3QZ</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>
+<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5vbu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5vbu OCA], [http://pdbe.org/5vbu PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5vbu RCSB], [http://www.ebi.ac.uk/pdbsum/5vbu PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5vbu ProSAT]</span></td></tr>
+</table>
+<div style="background-color:#fffaf0;">
+== Publication Abstract from PubMed ==
+Cytochrome P450 (P450, CYP) 21A2 is the major steroid 21-hydroxylase, converting progesterone to 11-deoxycorticosterone and 17alpha-hydroxyprogesterone (17alpha-OH-progesterone) to 11-deoxycortisol. More than 100 CYP21A2 variants give rise to congenital adrenal hyperplasia (CAH). We previously reported a structure of WT human P450 21A2 with bound progesterone and now present a structure bound to the other substrate (17alpha-OH-progesterone). We found that the 17alpha-OH-progesterone- and progesterone-bound complex structures are highly similar, with only some minor differences in surface loop regions. Twelve P450 21A2 variants associated with either salt-wasting or nonclassical forms of CAH were expressed, purified, and analyzed. The catalytic activities of these 12 variants ranged from 0.00009% to 30% of WT P450 21A2 and the extent of heme incorporation from 10% to 95% of the WT. Substrate dissociation constants (Ks) for four variants were 37-13,000-fold higher than for WT P450 21A2. Cytochrome b5, which augments several P450 activities, inhibited P450 21A2 activity. Similar to the WT enzyme, high noncompetitive intermolecular kinetic deuterium isotope effects (&gt;/= 5.5) were observed for all six P450 21A2 variants examined for 21-hydroxylation of 21-d3-progesterone, indicating that C-H bond breaking is a rate-limiting step over a 104-fold range of catalytic efficiency. Using UV-visible and CD spectroscopy, we found that P450 21A2 thermal stability assessed in bacterial cells and with purified enzymes differed among salt-wasting- and nonclassical-associated variants, but these differences did not correlate with catalytic activity. Our in-depth investigation of CAH-associated P450 21A2 variants reveals critical insight into the effects of disease-causing mutations on this important enzyme.
-Authors: Pallan, P.S., Egli, M.
+Functional analysis of human cytochrome P450 21A2 variants involved in congenital adrenal hyperplasia.,Wang C, Pallan PS, Zhang W, Lei L, Yoshimoto FK, Waterman MR, Egli M, Guengerich FP J Biol Chem. 2017 Jun 30;292(26):10767-10778. doi: 10.1074/jbc.M117.792465. Epub , 2017 May 24. PMID:28539365<ref>PMID:28539365</ref>
-Description: Crystal Structure of Human Cytochrome P450 21A2 Hydroxyprogesterone Complex
+From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
-[[Category: Unreleased Structures]]
+</div>
-[[Category: Pallan, P.S]]
+<div class="pdbe-citations 5vbu" style="background-color:#fffaf0;"></div>
+== References ==
+<references/>
+__TOC__
+</StructureSection>
 [[Category: Egli, M]]
+[[Category: Pallan, P S]]
+[[Category: Addison's disease]]
+[[Category: Adrenal steroidogenesis]]
+[[Category: Congenital adrenal hyperplasia]]
+[[Category: Hydroxyprogesterone]]
+[[Category: Kinetic isotope effect]]
+[[Category: Monooxygenase]]
+[[Category: Oxidoreductase]]
+[[Category: Steroid hydroxylation]]