<html><head><meta http-equiv="Content-Type" content="text/html; charset=utf-8"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">Hi Yeping,<div class=""><br class=""></div><div class=""> You have a difficult problem and Chimera is not going to be able to automatically produce the right fit, and neither am I. The preprint on the map you are fitting EMDB 22158 is here</div><div class=""><br class=""></div><div class=""><span class="Apple-tab-span" style="white-space:pre"> </span><a href="https://www.biorxiv.org/content/10.1101/2020.06.17.153486v2.full" class="">https://www.biorxiv.org/content/10.1101/2020.06.17.153486v2.full</a></div><div class=""><br class=""></div><div class="">They do not provide a PDB for the 22158 spike + antibody, although they do provide PDB 6xey for the related EMDB 22156 which is at higher resolution. Why is that? Probably because they are not confident of their antibody fit to 22158 or could not even make a trustworthy model of that antibody given the low resolution. What is the resolution? If you look at figure 13e where they show the local resolution, the antibody region has map resolution in the range 9.5 to 11.5 A. That is definitely not good enough to build an antibody model, nor even good enough to see the beta sheets of the antibody. By comparison look at the higher resolution EMDB 22156 and PDB 6xey they determined shown in figure 12 -- map resolution of around 4-6 A near the antibody, and they show their antibody model and density in figure 12e and you can see it is pretty reasonable.</div><div class=""><br class=""></div><div class=""> In summary, I don't know if your antibody atomic model is in the right conformation and you are trying to fit it into low ~10 Angstrom density. Chimera is not able to exhaustively sample configurations and tell you the best and even if it did it could very well be wrong with these uncertainties. Chimera is primarily used to interactively do fitting allowing you to understand whether a reliable fit is even possible.</div><div class=""><br class=""></div><div class=""> Good luck!</div><div class=""><br class=""></div><div class=""><span class="Apple-tab-span" style="white-space:pre"> </span>Tom</div><div class=""><br class=""></div><div class=""><br class=""></div><div class="">Fig 13 from paper.</div><div class=""><img apple-inline="yes" id="5B6BE0AC-88F1-4A6E-AB26-496A68505398" src="cid:8AFFBFF4-13D1-4C12-8203-96C5273E06A7" class=""><br class=""><div>Fig 12 from paper</div><div><br class=""></div><div><img apple-inline="yes" id="E5049AF0-0E47-4A05-9B2D-19D11C53CEF2" src="cid:18710A48-07E3-42C4-A139-A411F5748F95" class=""><br class=""><blockquote type="cite" class=""><div class="">On Aug 19, 2020, at 8:59 PM, sunyeping <<a href="mailto:sunyeping@aliyun.com" class="">sunyeping@aliyun.com</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div class="__aliyun_email_body_block"><div style="clear:both;" class="">Hi Tom,</div><div style="clear:both;" class=""><br class=""></div><div style="clear:both;" class="">Thank you for your suggestions and instructions. I have tried your method. Yes, the <span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class=""> "Optimize correlation" option in the "Fit in Map" tool can avoid large shit of the molecule. However, the problem with this method is that the final fitted result is determined by the initial position of the antibody molecule. I have no idea how the antibody binds the antigen, and my expectation is to find the correct binding positon of the antibody to the antigen by using the fitting tools of chimera. The only thing I know is that the CDRs face of the antibody should be placed towards the antigen. I have put the coordinations for the antigen (Ag, S.pdb) and the antibody (Ab, 4-8_H3-0175.pdb) at the following link:</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class=""> <a href="https://drive.google.com/drive/folders/16ELIDsUxiNwRGp9ely2SRn-zjAdHCudH?usp=sharing" class="">https://drive.google.com/drive/folders/16ELIDsUxiNwRGp9ely2SRn-zjAdHCudH?usp=sharing</a></span></div><div style="clear:both;" class=""><br class=""></div><div style="clear:both;" class="">Firstly, I fitted the Ag to the map for the ag-ab complex (emd_22158.map), then I put the antibody to target area of the map, and then used Fit in Map tool to fit it and get the complex coordinate fit-in-map_1.pdb. After that, I had another try. I put the Ab at the same initial position in the target area but rotated it by 180 degree around its long axis, then fit it, and get <span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class="">complex coordinate </span><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class="">fit-in-map_1.pdb. Both of the two </span> <span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class="">complex coordinate can also be found at the above link. In these two complex structures, the heavy chain (H) and light chain (L) of the Ab are totally opposite (a rotation of 180 degree). </span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class=""> Maybe you could try to do the fitting and find the most reasonable binding mode of the Ab? One clue is that the CDR3 of the H chain (residue 93-107) should usually be involved in contact with the antigen.</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class=""><br class=""></span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class="">Best regards,</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; text-align: start; text-indent: 0px; text-transform: none; widows: 2; background-color: rgb(255, 255, 255); float: none; display: inline;" class="">Yeping</span></div><blockquote style="margin-right:.0px;margin-top:.0px;margin-bottom:.0px;" class=""><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">------------------------------------------------------------------</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">From:Tom Goddard <<a href="mailto:goddard@sonic.net" class="">goddard@sonic.net</a>></span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">Sent At:2020 Aug. 20 (Thu.) 01:15</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">To:孙业平 <<a href="mailto:sunyeping@aliyun.com" class="">sunyeping@aliyun.com</a>></span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">Cc:chimera-users <<a href="mailto:chimera-users@cgl.ucsf.edu" class="">chimera-users@cgl.ucsf.edu</a>>; Greg Pintilie <<a href="mailto:gregdp@gmail.com" class="">gregdp@gmail.com</a>></span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class="">Subject:Re: [Chimera-users] Cannot dock molecule to targeted region?</span></div><div style="clear:both;" class=""><span style="font-family: Tahoma, Arial, STHeiti, SimSun; font-size: 14px;" class=""><br class=""></span></div>Hi Yeping,<div class=""><br class=""></div><div class=""> As Greg says Chimera is trying to move your antibody into higher density. Besides the masking he suggests, another thing you can do is just use the Fit in Map tool (not Fit to Segments), place your anti-body by hand in approximately the correct position using the "Active" checkbuttons in model panel and the mouse, and under options in the Fit in Map tool choose "Optimize correlation" instead of "Optimize overlap". The default optimize overlap means to move the molecule into the highest density, but optimizing correlation instead moves it to get the best agreement between the shape of the density. That option will be grayed out until you check "Use map simulated from atoms" since correlation needs a simulated map, and you should enter the approximate resolution of your map. I don't think Fit to Segments offers that option to optimize correlation. When you use Fit in Map it just does a local optimization so your initial placement and orientation of the antibody has to be approximately right.</div><div class=""><br class=""></div><div class=""> Tom</div><div class=""><br class=""></div><div class=""><span id="cid:__aliyun159789594465151061"><fit_in_map.png></span></div><div class=""><div class=""><br class=""><div class="">On Aug 19, 2020, at 5:13 AM, Greg Pintilie <<a class="" href="mailto:gregdp@gmail.com" target="_blank">gregdp@gmail.com</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div class="" style="overflow-wrap:break-word;"><div class=""><br class=""></div>Hi Yeping,<div class=""><br class=""></div><div class="">It seems that the part of the map you are fitting to has lower values, and the Fit-To-Segments procedure is moving the model (your antibody) towards higher values. To prevent this, there is an option in the Fit To Segments dialog ‘Mask map with region(s) to prevent large drifts’. If you enable this option you should not see the model move outside the region. You may have to press the Options button at the bottom of the dialog first to reveal this option if you don’t see it. Also make sure to try ‘Rotational search’ if the default/faster ‘Align principal axes’ doesn’t seem to find the right fit.</div><div class=""><br class=""></div><div class="">There is also the ‘fitmap’ command in Chimera which does a more global search, perhaps the Chimera team may have some advice on it in this scenario, but I think you have the right idea with segmenting first and then doing the fit - it’s a bit more work but you can target the search better with it.</div><div class=""><a class="" href="https://www.cgl.ucsf.edu/chimera/current/docs/UsersGuide/midas/fitmap.html" target="_blank">https://www.cgl.ucsf.edu/chimera/current/docs/UsersGuide/midas/fitmap.html</a></div><div class=""><br class=""></div><div class="">Greg</div><div class=""><br class=""><div class=""><br class=""><div class="">On Aug 19, 2020, at 1:53 AM, 孙业平 <<a class="" href="mailto:sunyeping@aliyun.com" target="_blank">sunyeping@aliyun.com</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;">Dear all,</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"><br class=""></span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"> I am trying to fit an antibody molecule to the cryo-EM density map for an antigen-antibody complex. The related files can be found at the following link</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"> <a class="" href="https://drive.google.com/drive/folders/16ELIDsUxiNwRGp9ely2SRn-zjAdHCudH?usp=sharing" target="_blank">https://drive.google.com/drive/folders/16ELIDsUxiNwRGp9ely2SRn-zjAdHCudH?usp=sharing</a>.</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;">I first segmented the map ("emd_22158.map") and get the segment map ("emd_22158.seg") in chimera. Then I selected the regions corresponding to the antibody and use the "Fit to segments" tool of chimera to fit the antibody molecule. The "image1.png" shows the inital antibody postion and the expected docking postion in the segmented complex density (indicated by the red arrow). However, the antibody molecule cannot be fitted into the selected target region after this operation, but moved to the position shown in "image2.png (only the target regions are shown). </span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"> I have tried to ungoup and regroup the target region to make its overall shape more simial to the antibody, and place the antibody into the region and ajust its orienation, but whatever I tried, the results are the same: the antibody will move to the position shown in "image2.png" after performing the <span class=" __aliyun_node_has_bgcolor" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;font-style:normal;font-variant-ligatures:normal;font-variant-caps:normal;font-weight:400;text-align:start;text-indent:.0px;text-transform:none;widows:2;background-color:#ffffff;float:none;display:inline;">"Fit to segments" action</span>.</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"> I wonder whether it is impossible to dock the antibody to the expected position with chimera or there is anything wrong with my operation. Could you help me with this issue? I will appreciate any help from you.</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;"><br class=""></span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;">Best regards,</span></div><div class="" style="clear:both;"><span class=" __aliyun_node_has_color" style="font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14.0px;">Yeping Sun</span></div>_______________________________________________<br class="">Chimera-users mailing list: <a class="" href="mailto:Chimera-users@cgl.ucsf.edu" target="_blank">Chimera-users@cgl.ucsf.edu</a><br class="">Manage subscription: <a class="" href="https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users" target="_blank">https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users</a><br class=""></div></div><br class=""></div></div>_______________________________________________<br class="">Chimera-users mailing list: <a class="" href="mailto:Chimera-users@cgl.ucsf.edu" target="_blank">Chimera-users@cgl.ucsf.edu</a><br class="">Manage subscription: <a class="" href="https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users" target="_blank">https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users</a><br class=""></div></div><br class=""></div></blockquote><div class=""><br class=""></div></div>_______________________________________________<br class="">Chimera-users mailing list: <a href="mailto:Chimera-users@cgl.ucsf.edu" class="">Chimera-users@cgl.ucsf.edu</a><br class="">Manage subscription: <a href="https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users" class="">https://plato.cgl.ucsf.edu/mailman/listinfo/chimera-users</a><br class=""></div></blockquote></div><br class=""></div></body></html>