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    On 13/03/2011 10:05 PM, Yulian Gavrilov wrote:
    <blockquote
      cite="mid:AANLkTi=4VX6J5LFpP4L74y7WykWidorhqtVzCDLGpTON@mail.gmail.com"
      type="cite">
      <div dir="ltr">Yes, I checked topol.top.  There are all isopeptide
        bonds ,that I want (according to atom contacts, angles, etc.)<br>
      </div>
    </blockquote>
    <br>
    Perhaps I've been labouring under a misapprehension. You've
    mentioned an isopeptide bond, which usually happens between a
    side-chain carboxylic acid (GLU,ASP)  and a side-chain amino acid
    (LYS), but also you've mentioned GLY, which only has a backbone
    carbonyl. My brain read "GLY" as "GLU". I would not expect pdb2gmx
    to cope correctly with terminating the glycine via specbond.dat.
    Please show us the [atoms] and [bonds] section for LYQ and GLYQ in
    your .top file.<br>
    <br>
    There were other points I made in an early email to which you've not
    responded, also.<br>
    <br>
    Mark<br>
    <br>
    <blockquote
      cite="mid:AANLkTi=4VX6J5LFpP4L74y7WykWidorhqtVzCDLGpTON@mail.gmail.com"
      type="cite">
      <div dir="ltr"><br>
        <div class="gmail_quote">2011/3/13 Mark Abraham <span dir="ltr">&lt;<a
              moz-do-not-send="true"
              href="mailto:Mark.Abraham@anu.edu.au">Mark.Abraham@anu.edu.au</a>&gt;</span><br>
          <blockquote class="gmail_quote" style="border-left: 1px solid
            rgb(204, 204, 204); margin: 0pt 0pt 0pt 0.8ex; padding-left:
            1ex;">
            <div text="#000000" bgcolor="#ffffff">
              <div class="im"> On 13/03/2011 9:53 PM, Yulian Gavrilov
                wrote:
                <blockquote type="cite">
                  <div dir="ltr">
                    <p style="margin-bottom: 0in;">Thank you!</p>
                    <p style="margin-bottom: 0in;"> </p>
                    <p style="margin-bottom: 0in;">I use the correct “O”
                      in Gly according to .rtp and I checked it with
                      vmd. There is really a new isopeptide bond. When
                      there is no bond, after minimization and
                      equilibration, Gly and Lys just close to each
                      other but they are not connected (in vmd). In my
                      case they are connected (in vmd, pymol). </p>
                  </div>
                </blockquote>
                <br>
              </div>
              None of these visualization programs read the topology in
              your .top file. They just make guesses based on the
              geometry of the atoms in the coordinate file. Anything you
              see that it guessed is irrelevant. Read your pdb2gmx
              output, and go and look at the topology to see what atoms
              have made a bond.<br>
              <font color="#888888"> <br>
                Mark</font>
              <div>
                <div class="h5"><br>
                  <br>
                  <blockquote type="cite">
                    <div dir="ltr">
                      <p style="margin-bottom: 0in;">When I look on
                        step...pdb, one these residues is exploded (it's
                        atoms are far from each other outside of the
                        water box).</p>
                      <br>
                      <br>
                      <div class="gmail_quote">2011/3/13 Mark Abraham <span
                          dir="ltr">&lt;<a moz-do-not-send="true"
                            href="mailto:Mark.Abraham@anu.edu.au"
                            target="_blank">Mark.Abraham@anu.edu.au</a>&gt;</span><br>
                        <blockquote class="gmail_quote"
                          style="border-left: 1px solid rgb(204, 204,
                          204); margin: 0pt 0pt 0pt 0.8ex; padding-left:
                          1ex;">
                          <div text="#000000" bgcolor="#ffffff">
                            <div> On 13/03/2011 8:55 PM, Yulian Gavrilov
                              wrote:
                              <blockquote type="cite">
                                <div dir="ltr">
                                  <p style="margin-bottom: 0in;">Dear
                                    gmx users,</p>
                                  <p style="margin-bottom: 0in;">I just
                                    started with gromacs.</p>
                                  <p style="margin-bottom: 0in;"> Can
                                    you help me to find my mistake? I
                                    already asked about it, but I did
                                    not understand what to do exactly in
                                    my case.</p>
                                  <p style="margin-bottom: 0in;">I try
                                    to run to add a new <b>isopeptide
                                      bone</b> to connect Lys and Gly
                                    (to make a tetramer of <b>ubiquitin</b>).

                                    I use AMBER99 force field, Gromacs
                                    4.0.5.</p>
                                  <p style="margin-bottom: 0in;">What I
                                    did:</p>
                                  <ol>
                                    <li>
                                      <p style="margin-bottom: 0in;">I
                                        changed names of residues
                                        according to AMBER rules (LYS to
                                        LYN etc.). </p>
                                    </li>
                                    <li>
                                      <p style="margin-bottom: 0in;">Added
                                        new type of residues to
                                        ffamber.rtp (LYN -&gt; LYQ and
                                        GLY -&gt; GLQ that are making a
                                        new isopeptide bond) and added a
                                        new line to specbond.dat (LYN NZ
                                        1 GLY C 1 0.13 LYQ GLQ) to make
                                        such a bond.</p>
                                    </li>
                                  </ol>
                                </div>
                              </blockquote>
                              <br>
                            </div>
                            IIRC, this creates a bond between the lysine
                            side-chain amine N and glycine *backbone*
                            carbonyl C. You must use the atom name for
                            the side-chain carbonyl carbon (see .rtp
                            entry for GLY). If you've done this wrong,
                            then specbond will probably not have made a
                            bond, because the backbone carbonyl C was
                            too far away. You should check your pdb2gmx
                            output carefully.
                            <div><br>
                              <br>
                              <blockquote type="cite">
                                <div dir="ltr">
                                  <ol>
                                    <li> <br>
                                    </li>
                                    <li>
                                      <p style="margin-bottom: 0in;">Added
                                        new bond type, angle type and
                                        dihedral type to
                                        ffamber99bon.itp</p>
                                    </li>
                                  </ol>
                                  <p style="margin-bottom: 0in;"><br>
                                  </p>
                                  <p style="margin-bottom: 0in;">After
                                    running of MD (I've got good
                                    minimization and equilibration – nvt
                                    and npt) I've got such an error:</p>
                                  <p style="margin-bottom: 0in;"><br>
                                  </p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">starting
                                      mdrun 'Protein in water'</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">600000
                                      steps, 1200.0 ps.</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">step
                                      94100, will finish Sun Mar 13
                                      08:15:56 2011</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">Step
                                      94124, time 188.248 (ps) LINCS
                                      WARNING</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">relative

                                      constraint deviation after LINCS:</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">rms
                                      0.000796, max 0.032792 (between
                                      atoms 2454 and 2456)</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">bonds
                                      that rotated more than 30 degrees:</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">atom
                                      1 atom 2 angle previous, current,
                                      constraint length</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">2454
                                      2457 35.6 0.1522 0.1545 0.1522</font></p>
                                  <p style="margin-bottom: 0in;"><br>
                                  </p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">And
                                      after it the same type of errors
                                      with another atoms:</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">2454
                                      2456 36.1 0.1106 0.1113 0.1090 <font
                                        size="3">--&gt; <b>Gly CA and
                                          HA1, HA2</b></font></font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">2454
                                      2455 40.5 0.1114 0.1103 0.1090</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">.....</font></p>
                                  <p style="margin-bottom: 0in;"> </p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">2454
                                      2456 90.0 0.1078 0.1479 0.1090</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">771
                                      773 48.5 0.1011 0.1012 0.1010</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">......</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">771
                                      773 39.8 0.1012 0.1005 0.1010 -<font
                                        size="3">-&gt; <b>Gly NZ and
                                          HZ1, HZ2</b></font></font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">771
                                      772 34.9 0.1012 0.1030 0.1010</font></p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">.......
                                    </font> </p>
                                  <p style="margin-bottom: 0in;"><font
                                      style="font-size: 9pt;" size="2">2454
                                      2457 102.1 0.1490
                                      38312886396780544.0000 0.1522</font></p>
                                  <p style="margin-bottom: 0in;"> <font
                                      style="font-size: 9pt;" size="2">2454
                                      2456 83.0 5.9313
                                      39290317874135040.0000 0.1090</font></p>
                                  <p style="margin-bottom: 0in;">.......</p>
                                  <p style="margin-bottom: 0in;">First
                                    errors are with atoms from the
                                    residues with <b>new isopeptide
                                      bond</b>. I suppose, that this
                                    bond is not good.</p>
                                </div>
                              </blockquote>
                              <br>
                            </div>
                            Seems like a reasonable hypothesis - but do
                            look at 2454 and 2456 as well. You have to
                            get out your trajectory and a visualization
                            package and see what is actually going
                            wrong. The warnings can be symptomatic of a
                            problem that started elsewhere.<br>
                            <br>
                            You say you've added new interaction types,
                            but I see no reason why you would need to.
                            It's chemically so similar to a backbone
                            peptide that you may as well keep things the
                            same and model it the same way. Regardless,
                            you should probably check that the
                            specbond.dat mechanism has created
                            interactions that make sense. Compare with a
                            normal peptide bond.<br>
                            <font color="#888888"> <br>
                              Mark</font>
                            <div><br>
                              <br>
                              <blockquote type="cite">
                                <div dir="ltr">
                                  <p style="margin-bottom: 0in;">Please
                                    can you advice me how yo make this
                                    isopeptide bond good?</p>
                                  <p style="margin-bottom: 0in;">Can I
                                    just remove this hydrogens?</p>
                                  <br>
                                  -- <br>
                                  <div dir="ltr">
                                    <p style="margin-bottom: 0in;">Sincerely,</p>
                                    <p style="margin-bottom: 0in;">Yulian
                                      Gavrilov </p>
                                  </div>
                                  <br>
                                </div>
                              </blockquote>
                              <br>
                            </div>
                          </div>
                          <br>
                          --<br>
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                        </blockquote>
                      </div>
                      <br>
                      <br clear="all">
                      <br>
                      -- <br>
                      <div dir="ltr">
                        <p style="margin-bottom: 0in;"> Sincerely,</p>
                        <p style="margin-bottom: 0in;">Yulian Gavrilov </p>
                      </div>
                      <br>
                    </div>
                  </blockquote>
                  <br>
                </div>
              </div>
            </div>
            <br>
            --<br>
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        <br>
        -- <br>
        <div dir="ltr">
          <p style="margin-bottom: 0in;">
            Sincerely,</p>
          <p style="margin-bottom: 0in;">Yulian Gavrilov </p>
        </div>
        <br>
      </div>
    </blockquote>
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