The X-ray menu (XRAY)

Introduction.

WARNING: At present the density fitting routines only work for orthogonal
maps with equal stepsize in all three directions.

The X-ray menu holds the options mainly meant for crystallography. At present there are commands to convolute desity with atoms, to use symmetry matrices to objects or parts of the soup, and to do some real space refinement. The real space refinement options assume that you already used the map menu to read an electron density map. The symmetry options assume that you already obtained symmetry matrices.

Convolution of atoms with density

In the electron density convolution options atoms are represented by a sphere with a bell shaped radial density distribution. See:

R. Voorintholt, M.T. Kosters, G. Vegter, G. Vriend, W.G.J. Hol, A very fast program for visualizing protein surfaces, channels and cavities. (1989) J. Mol. Graph. pp 243-245.

The convolution is the sum of all products rho(i)*atm(i) in which i runs over the volume of the atom, atm(i) is the value of the radial bell-shaped atom density at position i, and rho(i) is the electron density map value at that same position in space.

The results of the density convolution options are NOT on any absolute scale. You can only compare values within one run of this option. If you want to compare values between different maps, then these maps should have the same statistics, e.g. the same average density, and the same sigma (=spread in density).

Convoluting electron density with all atoms (EVADEN)

The option EVADEN will cause WHAT IF to prompt you for a residue range. It will then convolute all atoms in this range with the density found in the present default map. The density under each atom will be listed as well as the summed density per residue. You will be asked if you want to store the results in a table. If you answer with YES, you will as usual (see the chapter on tables) be prompted for the table number and the table name.

Convoluting electron density with alpha carbons (EVACAS)

The option EVACAS will cause WHAT IF to prompt you for a residue range. It will then convolute all alpha carbons in this range with the density found in the present default map. The density under each alpha carbon will be listed. You will be asked if you want to store the results in a table. If you answer with YES, you will as usual (see the chapter on tables) be prompted for the table number and the table name.

Real space refinement

Side chain optimization (RSRSID)

The RSRSID option will prompt you for a residue range. It will try to optimize the convolution between the density and bell shaped side chain atoms for the residues in this range. You will also be prompted for the allowed Van der Waals radius overlap. The suggested value is 0.25 Angstrom. This number indicates how far two atoms are allowed to penetrate each other. You will be asked to give the number of steps per chi-angle. The suggested value is 2. This number tells WHAT IF how many steps to try in either direction for every free rotatable side chain torsion angle. Choosing 6 means that for lysine 13*13*13*13 density convolutions (13 values for each of 4 free rotatable bonds) have to be done. The last value you are prompted for is the angle per step. In early stages of building you might give 20 degrees or so. In later stages a value less than 5 degrees is suggested.

Density fit optimization (RSRRES)

The command RSRRES will cause WHAT IF to prompt you for a residue range. For all residues in this range it will move them around in small translational steps (maximal step per call to this option is 0.4 Angstrom) to search for the optimal convolution between the atom positions and the electron density. This is a rigid body motion.

Density fit optimization (RSRBB)

The option RSRBB does the same as the option RSRRES. The difference is that RSRBB only tries to optimize the backbone density fit. It will however apply the translation used for the backbone to the sidechain too.

Graphical use of symmetry matrices

The graphical expansion of MOL-items or the FBRT or MOVITM object through the usage of symmetry matrices is probably only useful for crystallographers. Also, it relies heavily on some Evans and Sutherland hardware features Todate the option only works on E and S machines, and it is not likely that it will run equally well on other machines once the conversion is done.

Apply symmetry matrices to a MOL-object (EXPSYM)

EXPSYM is used to apply symmetry matrices to a MOL-object. These matrices can be crystallographic or non-crystallographic. You can give `normal` symmetry matrices or real space symmetry matrices. You will be prompted for the number of a MOL-item. All presently active symmetry matrices (see the chapter on symmetry matrices) will be applied to this MOL-object. The symmetry expanded object is stored in MOL-object 8. That means that MOL-object 8 can not be used as input for this option.

The flow of events to use this option is:

1) Get the symmetry matrices (see the SYM menu).

2) Create a MOL-item in a MOL-object, but not in MOL-8.

3) Use the EXPSYM option.

Step 2 and 3 can be interchanged. It is not possible to undo the EXPSYM option. It will stay active.

Applying symmetry matrices to fbrt (FBRSYM)

FBRSYM can be used to move part of the soup as a FBRT object (see FBRT in the chapter on graphics options) while at the same time moving the symmetry related parts of the soup in accordance with the symmetry matrices. These matrices can be crystallographic or non-crystallographic. You can give `normal` symmetry matrices or real space symmetry matrices. Remember that the MOVITM object is also stored in FBRT.

The flow of events to use thsi option is:

1) Get the symmetry matrices (see the SYM menu).

2) Uset the FBRSYM option.

3) Pass control to the graphics screen/window.

4) Use the FBRT option.

The symmetry related parts of the soup are automatically put in MOL-object 0 (zero) by WHAT IF. FBRSYM can not be switched off. It works on every next FBRT or MOVITM option. If you click YES at the graphics screen menu the present matrix that you applied to the FBRT/MOVITM object will be displayed. This is always a real space matrix.

Other X-ray related options

Other Xray related options are found in the menus: MAP, MAPEDT, CHKMDF, WATER, REFINE, etc..

Flipping peptide planes (PEPFLP)

If the either the density or a WHAT IF checking option suggests that a peptide plane needs flipping, you can use this option to do so. Be aware however, that backbone bond angles are no longer OK after such an operation, and subsequent normalisation with REFI in the REFINE menu seems required if you still want to call your molecule a protein afterwards.

Reversing the backbone of a residue range (RNGFLP)

Sometimes, upon density fitting in poor density, one builds a stretch of residues in the wrong direction. The RNGFLP option can invert the chain direction of a range of residues, without the need to refit everything afterwards. This option is likely to perform better for beta-like structures than for highly curved secondary structure elements. The range will be returned as poly-alanine.