1cbw: Difference between revisions

Revision as of 13:04, 21 February 2008

BOVINE CHYMOTRYPSIN COMPLEXED TO BPTI

OverviewOverview

The crystal structures of the inhibitor domain of Alzheimer's amyloid beta-protein precursor (APPI) complexed to bovine chymotrypsin (C-APPI) and trypsin (T-APPI) and basic pancreatic trypsin inhibitor (BPTI) bound to chymotrypsin (C-BPTI) have been solved and analyzed at 2.1 A, 1.8 A, and 2.6 A resolution, respectively. APPI and BPTI belong to the Kunitz family of inhibitors, which is characterized by a distinctive tertiary fold with three conserved disulfide bonds. At the specificity-determining site of these inhibitors (P1), residue 15(I)4 is an arginine in APPI and a lysine in BPTI, residue types that are counter to the chymotryptic hydrophobic specificity. In the chymotrypsin complexes, the Arg and Lys P1 side chains of the inhibitors adopt conformations that bend away from the bottom of the binding pocket to interact productively with elements of the binding pocket other than those observed for specificity-matched P1 side chains. The stereochemistry of the nucleophilic hydroxyl of Ser 195 in chymotrypsin relative to the scissile P1 bond of the inhibitors is identical to that observed for these groups in the trypsin-APPI complex, where Arg 15(I) is an optimal side chain for tryptic specificity. To further evaluate the diversity of sequences that can be accommodated by one of these inhibitors, APPI, we used phage display to randomly mutate residues 11, 13, 15, 17, and 19, which are major binding determinants. Inhibitors variants were selected that bound to either trypsin or chymotrypsin. As expected, trypsin specificity was principally directed by having a basic side chain at P1 (position 15); however, the P1 residues that were selected for chymotrypsin binding were His and Asn, rather than the expected large hydrophobic types. This can be rationalized by modeling these hydrophilic side chains to have similar H-bonding interactions to those observed in the structures of the described complexes. The specificity, or lack thereof, for the other individual subsites is discussed in the context of the "allowed" residues determined from a phage display mutagenesis selection experiment.

About this StructureAbout this Structure

1CBW is a Protein complex structure of sequences from Bos taurus with as ligand. Active as Chymotrypsin, with EC number 3.4.21.1 Full crystallographic information is available from OCA.

ReferenceReference

Crystal structures of bovine chymotrypsin and trypsin complexed to the inhibitor domain of Alzheimer's amyloid beta-protein precursor (APPI) and basic pancreatic trypsin inhibitor (BPTI): engineering of inhibitors with altered specificities., Scheidig AJ, Hynes TR, Pelletier LA, Wells JA, Kossiakoff AA, Protein Sci. 1997 Sep;6(9):1806-24. PMID:9300481

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-[[Image:1cbw.jpg|left|200px]]<br /><applet load="1cbw" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1cbw.jpg|left|200px]]<br /><applet load="1cbw" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1cbw, resolution 2.6&Aring;" />
 '''BOVINE CHYMOTRYPSIN COMPLEXED TO BPTI'''<br />
 ==Overview==
-The crystal structures of the inhibitor domain of Alzheimer's amyloid, beta-protein precursor (APPI) complexed to bovine chymotrypsin (C-APPI), and trypsin (T-APPI) and basic pancreatic trypsin inhibitor (BPTI) bound, to chymotrypsin (C-BPTI) have been solved and analyzed at 2.1 A, 1.8 A, and 2.6 A resolution, respectively. APPI and BPTI belong to the Kunitz, family of inhibitors, which is characterized by a distinctive tertiary, fold with three conserved disulfide bonds. At the specificity-determining, site of these inhibitors (P1), residue 15(I)4 is an arginine in APPI and a, lysine in BPTI, residue types that are counter to the chymotryptic, hydrophobic specificity. In the chymotrypsin complexes, the Arg and Lys P1, side chains of the inhibitors adopt conformations that bend away from the, bottom of the binding pocket to interact productively with elements of the, binding pocket other than those observed for specificity-matched P1 side, chains. The stereochemistry of the nucleophilic hydroxyl of Ser 195 in, chymotrypsin relative to the scissile P1 bond of the inhibitors is, identical to that observed for these groups in the trypsin-APPI complex, where Arg 15(I) is an optimal side chain for tryptic specificity. To, further evaluate the diversity of sequences that can be accommodated by, one of these inhibitors, APPI, we used phage display to randomly mutate, residues 11, 13, 15, 17, and 19, which are major binding determinants., Inhibitors variants were selected that bound to either trypsin or, chymotrypsin. As expected, trypsin specificity was principally directed by, having a basic side chain at P1 (position 15); however, the P1 residues, that were selected for chymotrypsin binding were His and Asn, rather than, the expected large hydrophobic types. This can be rationalized by modeling, these hydrophilic side chains to have similar H-bonding interactions to, those observed in the structures of the described complexes. The, specificity, or lack thereof, for the other individual subsites is, discussed in the context of the "allowed" residues determined from a phage, display mutagenesis selection experiment.
+The crystal structures of the inhibitor domain of Alzheimer's amyloid beta-protein precursor (APPI) complexed to bovine chymotrypsin (C-APPI) and trypsin (T-APPI) and basic pancreatic trypsin inhibitor (BPTI) bound to chymotrypsin (C-BPTI) have been solved and analyzed at 2.1 A, 1.8 A, and 2.6 A resolution, respectively. APPI and BPTI belong to the Kunitz family of inhibitors, which is characterized by a distinctive tertiary fold with three conserved disulfide bonds. At the specificity-determining site of these inhibitors (P1), residue 15(I)4 is an arginine in APPI and a lysine in BPTI, residue types that are counter to the chymotryptic hydrophobic specificity. In the chymotrypsin complexes, the Arg and Lys P1 side chains of the inhibitors adopt conformations that bend away from the bottom of the binding pocket to interact productively with elements of the binding pocket other than those observed for specificity-matched P1 side chains. The stereochemistry of the nucleophilic hydroxyl of Ser 195 in chymotrypsin relative to the scissile P1 bond of the inhibitors is identical to that observed for these groups in the trypsin-APPI complex, where Arg 15(I) is an optimal side chain for tryptic specificity. To further evaluate the diversity of sequences that can be accommodated by one of these inhibitors, APPI, we used phage display to randomly mutate residues 11, 13, 15, 17, and 19, which are major binding determinants. Inhibitors variants were selected that bound to either trypsin or chymotrypsin. As expected, trypsin specificity was principally directed by having a basic side chain at P1 (position 15); however, the P1 residues that were selected for chymotrypsin binding were His and Asn, rather than the expected large hydrophobic types. This can be rationalized by modeling these hydrophilic side chains to have similar H-bonding interactions to those observed in the structures of the described complexes. The specificity, or lack thereof, for the other individual subsites is discussed in the context of the "allowed" residues determined from a phage display mutagenesis selection experiment.
 ==About this Structure==
-CBW is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bos_taurus Bos taurus] with SO4 as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Chymotrypsin Chymotrypsin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.1 3.4.21.1] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1CBW OCA].
+CBW is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bos_taurus Bos taurus] with <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Chymotrypsin Chymotrypsin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.1 3.4.21.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1CBW OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Chymotrypsin]]
 [[Category: Protein complex]]
-[[Category: Hynes, T.R.]]
+[[Category: Hynes, T R.]]
-[[Category: Kossiakoff, A.A.]]
+[[Category: Kossiakoff, A A.]]
-[[Category: Scheidig, A.J.]]
+[[Category: Scheidig, A J.]]
 [[Category: SO4]]
 [[Category: complex (serine protease/inhibitor)]]
@@ Line 23: / Line 23: @@
 [[Category: serine protease]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 12:19:44 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:04:27 2008''