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Set up Membrane Bilayer system by CHARMM-GUI

Step 00 - PDB Info

  • Download 4QTN from OPM database

Step 01 - Select Model/Chain, Manipulate PDB, Generate PDB and Orient Molecule

  • Select Chain A, B, and C, with residue ID 2-236. #Also add NNR if to make PnuC_3NR system.
  • Use the PDB orientation.
  • If NNR cannot be recognized by the system, that means atom numbering system is not the same as PDB record. You may need to download NNR from RCSB PDB and insert/replace the one in your own model.

Step 02 - Determine the system size

#Calculate Cross-Sectional Area

#System Size Determination Options:

  • Heterogeneous Lipid
  • 1. Box Type: Rectangular
  • 2. Length of Z based on: Water thickness (Minimum water height on top and bottom of the system)
  • 3. Length of XY based on: Ratios of lipid components, POPE:POPG = 3:1
  • Further optimize by check Numbers of lipid components, and change numbers of lipids to eliminate any errors
  • POPE: 143 139 Upperleaflet Lowerleaflet
  • POPG: 48 46 Upperleaflet Lowerleaflet

#Results:

  • Box Type Rectangle
  • Crystal Type TETRAGONAL
  • System Size

A 120.581738 Dimension along the A (X) axis B 120.581738 Dimension along the B (Y) axis C 104.427824 Dimension along the C (Z) axis

-Crystal Angle

Alpha 90.0 Angle between the axis B and C Beta 90.0 Angle between the axis A and C Gamma 90.0 Angle between the axis A and B

#of Lipids

on Top 191 on Bottom 185 Z Center -2.77291175 Center of the system along the Z axis

Step 03 - Build Components

  • System Building Options:

  • Replacement method Build system using replacement method Check lipid ring (and protein surface) penetration

  • Component Building Options:

  • Include Ions : 0.15 M NaCl (ion concentration)

  • Ion Placing Method: Monte-Carlo