2N8R |

Productive complex between MMP-12 and synthetic triple-helical collagen, revealed through paramagnetic NMR

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.4 mM [U-99% 15N] MMP, 0.6 mM TOAC labelled in P5 position THP	90% H2O/10% D2O		6.6	ambient	299
2	2D 1H-15N HSQC	0.25 mM [U-100% 12C; U-100% 15N; U-100% 2H; U-100% 13CH3] MMP, 0.38 mM TOAC labelled in P8' position THP	90% H2O/10% D2O		6.6	ambient	299
3	2D 1H-13C HSQC	0.25 mM [U-100% 12C; U-100% 15N; U-100% 2H; U-100% 13CH3] MMP, 0.38 mM TOAC labelled in P8' position THP	90% H2O/10% D2O		6.6	ambient	299

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	Avance	800

NMR Refinement
Method	Details	Software
Rigid-body docking, Conformer selection	The Collagen triple helix repeat peptide chain is homology-modeled from 4AUO. The Macrophage metalloelastase enzyme starting structure is 2poj.	TOPSPIN

NMR Ensemble Information
Conformer Selection Criteria	back calculated data agree with experimental NOESY spectrum
Conformers Calculated Total Number	7500
Conformers Submitted Total Number	1
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TOPSPIN		Bruker Biospin
2	processing	TOPSPIN		Bruker Biospin
3	chemical shift assignment	Analysis		CCPN
4	peak picking	Analysis		CCPN
5	structure solution	HADDOCK	2.1	Alexandre Bonvin
6	structure solution	q_test.py		Stephen H. Prior
7	refinement	GROMOS		van Gunsteren and Berendsen
8	refinement	HADDOCK	2.1	Alexandre Bonvin
9	refinement	q_test.py		Stephen H. Prior

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM157729.