ࡱ> ;=D,-./0123456789:<ܥhW  e"fU]3NlNllNlNNNNNONONONONONOjO4NOT]OOOOOPPPQc6R6R6R,bRdSfTTX9U-TNPODPPPTzQlNlNOOzQzQzQPlN8ONOQJbN8N^lNlNlNlNPQzQYzQ1.1 Basic features The moment you open the file PGCS01.WK4, a screen like in fig 1.1 appears. Notice that the usual menus of Lotus123 are replaced by PCGS menus (to return to the Lotus123 menu click the mouse on the last option in PGCS menu [Reset]). By clicking the mouse on the various sheets title you can view and/or modify their contents.  Fig. 1.1 Appearance of the first screen of PGCS.  1.2 Sheets Contents 1.2.1 Title Sheet This is the sheet you will access by default every time you open PGCS. It contains the title legend (fig. 1.1). At lower sheet areas, the title sheet also contains a results summary area (averages of dihedral and helix parameters) and a Ramachandran plot area. These areas are updated every time a new calculation of dihedral angles is run. The Ramachandran plot and results summary are easily accessed from anywhere in PGCS through the menu item [Summary]. 1.2.2 Workshop This sheet contains all the formulae and transformations necessary for calculating the dihedral angles and the helix parameters. It should not be modified by the user. A listing of the formulae in this sheet is given in Appendix II. 1.2.3 Dihedral Angles (output of dihedral angles calculation) The results of dihedral angles and helix parameters will be located in this sheet. 1.2.3 Geometry Data (input sheet) In this sheet the user is going to paste the geometry specifications in PDB format. 1.2.4 Dist. Matrix (output of distance matrix calculation) The distance matrix will show in this sheet if PGCS is requested to compute it. 1.2.5 Macros and Code This sheet contains all the macros and routines controlling the calculations and outputs. It should not be modified by the user. A listing of macros and code is given in Appendix I. 2. Tutorial I: Backbone Dihedrals and Parameters Click the mouse on [Geometry Data] access header (see fig 1.1). You will jump to the Geometry Data sheet and should have a screen similar to fig. 1.2. The sheet contains sample data for the purpose of this tutorial. Later you will erase this data and enter your own in the same format (PDB format). Now save your file under a new name, keeping the original PCGS copy unmodified as a backup. To save the file you must revert to the standard Lotus123 menu by using the PCGS menu [Reset], and save as usual using the [File] option. Fig. 2.1 Geometry Data Sheet   Get back to the PCGS Menu After you reverted to the standard Lotus123 menu, you may return back to the PCGS menu at any time using the key combination: Cntrl + P Run the first calculation  Notice that a help item appears at the very top of the screen each time the mouse move to an option in any menu (or sub-menu). 1. Click the mouse on the [Setup] menu. (See fig. 2.1) A list of options appear. We will not use the first option [Clear previous bond lengths and angles], as this is our first calculation. 2. Click on the second option [Sort data for backbone Dihedrals]. The data will be sorted in the order necessary to compute the backbone dihedrals. All side chain atoms as well as hydrogens are sorted out of the active part of the data (at the bottom of the data list). At this stage, PGCS automatically compute bond angles (no the dihedrals yet) and bond distances in sequence as listed after the sort command. The sequential bond angles and distances are listed to the right of the geometry data as shown in fig. 2.2. For example, the first bond distance N285-C(286 is 14640 , the distance C287-C(286 is 1.5249 and the angle C287-C(-N is 110.7. Fig 2.2 Appearance of the sheet [Geometry Data] after the [Setup/Sort data for backbone dihedrals] option.  3. Click the mouse again on the [Setup] menu and then on the sixth option [Number of cycles [Dihedrals]]. You will get a dialog box as in fig. 2.3. The recommended number of dihedral is the maximum number of backbone dihedrals that can be computed from the data in the [Geometry Data] sheet. The user may want all the backbone dihedrals (all the (, (, an ( angles. Or he/she may wish to compute, say, only the first 10 dihedral angles. In the first case the user enters the number of dihedrals recommended by the program, 37 in this tutorial, and 10 in the second case. If the user enters a number greater than the recommended one, the next macro will execute compute all the dihedral and continue in the same order for the remainder of the data. Any angle computed after all the defined backbone dihedral have been computed does not have a standard definition and cannot be correctly labeled as (, (, or (. Thus, the user should enter a number equal to or smaller than the recommended number of dihedrals, but never exceed that number. Fill in the number of dihedrals and Click OK. You will jump to the Dihedral Angles sheet. Now you are ready to compute the dihedrals. Fig. 2.3 Dialog box from [Setup/Number of Cycles [Dihedrals]]  4. Click on the menu [Dihedral Angles]. A list of options will appear as in fig. 2.4. The first option [Compute ALL dihedrals parameters and labels] is the one users will utilize most often. Clicking on this option will result in labeling as well as calculating all dihedral angles along the backbone of the peptide ((, (, and (), as well as the ( angle, the height h per turn, and the number n of residues per turn ((, h, and n are only meaningful in case of helices). The options [Labels] and [Compute dihedrals and parameters] breakdown the first option [Compute ALL dihedrals and parameters] into two parts: [Labels] will only read the data in the Geometry data sheet and produces a list of labeled dihedrals. [Compute dihedrals ] will compute the dihedrals without labeling them. The option [Single dihedral angle list] provides the freedom to the user to sort the Geometry data in any order that he/she would prefer (not necessary the sorting options defined in PGCS), and then compute the dihedrals in sequence starting from top to the bottom of the list. Since this list may have another definition than (, (, (, or (, the output is printed in the unlabelled column on the left of the column ( in the output (see fig.2.5). The last option [One single dihedral angle] is similar to option (c), but computes one dihedral angle each time, until the number of dihedrals specified by the user (in dialog box - fig. 2.3) is reached. Fig. 2.4 Menu [Dihedral Angles]  The user should obtain a list which looks like the one in fig. 2.5 by now. Fig 2.5 Output of the dihedral angles calculation.       Now if you wish to access average values for dihedrals and parameters, or if you want to access the Ramachandran plot, click the menu [Summary] then the appropriate option. A Ramachandran plot and averages corresponding to the results of fig.25 are shown in figure 2.6 and 2.7 respectively. Fig. 2.6 Ramachandran plot corresponding to the results of dihedral angle calculations shown in fig. 2.5.  Fig. 2.7 Summary (average values) corresponding to the results of dihedral angle calculations shown in fig. 2.5.  2. Tutorial II: Hydrogen bonds  Before next Run Clear the previous bond angles and distances using the menu sequence [Setup]/[Clear previous bond lengths and angles]. Always do this step. 1. Click on the menu [Setup]/[Sort data for H-N-C-Ha torsion angles]. Cartesian Coordinates: PGCS menu bar Button to revert to the PGCS menu Button to revert to the standard Lotus 123 menu PGCS sheets titles & accsess headers Help item Amino acid number in the polypeptide sequence Amino acid to which the last Nitrogen belongs Amino acid to which the first Nitrogen belongs Atom numbers of the N, C(, and C respectively belonging to the amino acid on the left. The last number is that of the N belonging to the next amino acid in the sequence, i.e. the amino acid on the right. 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߿ dsNNq ??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ߿ sNNNN߯ ?????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????  4NNA ????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? 4A ????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? 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T:Im>)d# ) )"Systemn- "--   \ Arial-2  IAA #Times New Roman)--2 J (Atom Numbers    -2 `1AA #-)2 2 Dihedral Angles   Arial-- 2 Pa%N2 d`y%CA 2 x%C 2 1%NSymbol-- 2 Ny%y 2 x%f 2 %w 2  I%d- 2 hH%h 2 %n- 2 #\1 Arial--2 #*(#I4ASP  Arial-2 %J\2852 %b\2862 %z\2872 %\296-2 ##4THR - 2 #\2 Arial- 2 #B\-179.0642 #\-65.4572 #\-177.164-2 #\-223.4012 #e\1.37632 #\-1.61 2 2\2-2 2*(2ICTHR -2 4J\2962 4b\2972 4z\2982 4\305-2 22CALA - 2 2\3- 2 2I\-31.0662 2\-64.1992 2\-178.250-2 2\107.5402 2e\1.40562 2\3.35 2 A\3-2 A*(AIRALA -2 CJ\3052 Cb\3062 Cz\3072 C\311-2 AARSER - 2 A\4- 2 AI\-43.4022 A\-68.4692 A\-176.554-2 A\113.3052 Ae\1.34972 A\3.18 2 P\4-2 P*(PIaSER -2 RJ\3112 Rb\3122 Rz\3132 R\319-2 PPaASP - 2 P\5- 2 PI\-40.9952 P\-63.7952 P\-178.366-2 P\110.3712 Pe\1.37402 P\3.26 2 _\5-2 _*(_IpASP -2 aJ\3192 ab\3202 az\3212 a\328-2 __pALA - 2 _\6- 2 _I\-41.3172 _\-67.6662 _\179.729-2 _\112.0372 _e\1.35872 _\3.21 2 n\6-2 n*(nIALA -2 pJ\3282 pb\3292 pz\3302 p\334-2 nnALA - 2 n\7- 2 nI\-35.2572 n\-66.0902 n\176.448-2 n\109.2772 ne\1.38372 n\3.29 2 }\7-2 }*(}IALA -2 J\3342 b\3352 z\3362 \340-2 }}ALA - 2 }\8- 2 }I\-44.9752 }\-66.1152 }\-179.376-2 }\112.9442 }e\1.35342 }\3.19 2 \8-2 *(IALA -2 J\3402 b\3412 z\3422 \346-2 ALA - 2 \9- 2 I\-37.4152 \-62.1552 \-177.010-2 \108.6252 e\1.39202 \3.31 2 \9-2 *(IALA -2 J\3462 b\3472 z\3482 \352-2 ALA -2 \10- 2 I\-43.8362 \-60.4152 \176.638-2 \110.2022 e\1.37782 \3.272 \10-2 *(IALA -2 J\3522 b\3532 z\3542 \358-2 ALA -2 \11- 2 I\-46.4552 \-66.6522 \177.520-2 \113.8972 e\1.34732 \3.162 \11-2 *(IALA -2 J\3582 b\3592 z\3602 \364-2 LEU -2 \12- 2 I\-42.7952 \-67.3492 \-178.426-2 \112.5232 e\1.35542 \3.202 \12-2 *(ILEU -2 J\3642 b\3652 z\3662 \373-2 THR -2 \13- 2 I\-43.5872 \-61.3592 \-177.471-2 \110.4442 e\1.37502 \3.262 \13-2 *(ITHR -2 J\3732 b\3742 z\3752 \382-2 ALA -2 \14- 2 I\-42.6942 \-62.3082 \-178.267-2 \110.4522 e\1.37422 \3.262 \14-2 *(IALA -2 J\3822 b\3832 z\3842 \388-2 ALA -2 \15- 2 I\-42.7872 \-64.2652 \-179.032-2 \111.2312 e\1.36662 \3.242 \15-2 *(IALA -2 J\3882 b\3892 z\3902 \394-2 ASN -2 \16- 2 I\-39.5412 \-69.6932 \-178.581-2 \112.1962 e\1.35672 \3.212 \16-2 *(IASN -2 J\3942 b\3952 z\3962 \405-2 ALA -2 \17- 2 I\-38.3732 \-65.1022 \177.833-2 \109.9272 e\1.37742 \3.272 \17-2 *(I$ALA -2 J\4052 b\4062 z\4072 \411-2 $LYS -2 \18- 2 I\-40.7552 \-62.8572 \173.768-2 \109.9452 e\1.37832 \3.272 "\18-2 "*("I3LYS -2 $J\4112 $b\4122 $z\4132 $\424-2 ""3ALA -2 "\19- 2 "I\-40.8442 "\-61.5452 "\-177.952-2 "\109.5222 "e\1.38302 "\3.292 1\19-2 1*(1IBALA -2 3J\4242 3b\4252 3z\4262 3\430-2 11BALA -2 1\20- 2 1I\-43.4042 1\-61.5682 1\-176.609-2 1\110.4512 1e\1.37482 1\3.262 @\20-2 @*(@IQALA -2 BJ\4302 Bb\4312 Bz\4322 B\436-2 @@QALA -2 @\21- 2 @I\-36.8912 @\-68.3832 @\179.290-2 @\110.6552 @e\1.36972 @\3.252 N\21-2 N*(OI`ALA -2 PJ\4362 Pb\4372 Pz\4382 P\442-2 NO`GLU -2 N\22- 2 NI\-35.2382 N\-66.6892 N\-177.339-2 N\109.4812 Ne\1.38142 N\3.292 ]\22-2 ]*(^IoGLU -2 _J\4422 _b\4432 _z\4442 _\452-2 ]^oLEU -2 ]\23- 2 ]I\-40.6362 ]\-65.1882 ]\-178.594-2 ]\110.7632 ]e\1.36992 ]\3.252 l\23-2 l*(mI~LEU -2 nJ\4522 nb\4532 nz\4542 n\461-2 lm~THR -2 l\24- 2 lI\-42.6582 l\-60.2732 l\176.448-2 l\109.7232 le\1.38212 l\3.282 {\24-2 {*(|ITHR -2 }J\4612 }b\4622 }z\4632 }\470-2 {|ALA -2 {\25- 2 {I\-46.4362 {\-63.9722 {\179.128-2 {\112.6682 {e\1.35682 {\3.202 \25-2 *(IALA -2 J\4702 b\4712 z\4722 \476-2 ALA -2 \26- 2 I\-43.2392 \-61.7622 \-177.018-2 \110.4592 e\1.37462 \3.262 \26-2 *(IALA -2 J\4762 b\4772 z\4782 \482-2 ASN -2 \27- 2 I\-38.0522 \-69.6922 \-176.845-2 \111.6112 e\1.36122 \3.232 \27-2 *(IASN -2 J\4822 b\4832 z\4842 \493-2 ALA -2 \28- 2 I\-42.8932 \-65.0022 \178.071-2 \111.5692 e\1.36362 \3.232 \28-2 *(IALA -2 J\4932 b\4942 z\4952 \499-2 ALA -2 \29- 2 I\-41.1072 \-62.6412 \176.678-2 \109.9932 e\1.37802 \3.272 \29-2 *(IALA -2 J\4992 b\5002 z\5012 \505-2 ALA -2 \30- 2 I\-41.5112 \-62.8522 \-178.327-2 \110.2142 e\1.37592 \3.272 \30-2 *(IALA -2 J\5052 b\5062 z\5072 \511-2 ALA -2 \31- 2 I\-40.2672 \-61.9972 \-178.058-2 \109.4802 e\1.38322 \3.292 \31-2 *(IALA -2 J\5112 b\5122 z\5132 \517-2 ALA -2 \32- 2 I\-47.1942 \-60.7362 \179.600-2 \111.6722 e\1.36612 \3.222 \32-2 *(IALA -2 J\5172 b\5182 z\5192 \523-2 ALA -2 \33- 2 I\-45.1792 \-66.9642 \-178.324-2 \113.4282 e\1.34992 \3.172 \33-2 *(IALA -2 J\5232 b\5242 z\5252 \529-2 ALA -2 \34- 2 I\-37.1722 \-66.2722 \-179.371-2 \109.9512 e\1.37682 \3.272 \34-2 *(I"ALA -2 J\5292 b\5302 z\5312 \535-2 "THR -2 \35- 2 I\-38.3412 \-62.1562 \178.352-2 \108.9082 e\1.38892 \3.312 \35-2 *( I1THR -2 "J\5352 "b\5362 "z\5372 "\544-2  1ALA -2 \36- 2 I\-39.5512 \-70.1512 \177.606-2 \112.4042 e\1.35512 \3.202 /\36-2 /*(/I@ALA -2 1J\5442 1b\5452 1z\5462 1\550-2 //@ARG -2 /\37- 2 /I\-23.2992 /\-99.9692 /\178.639-2 /\124.0172 /e\1.30142 /\2.902 >\37-2 >*(>IOARG -2 @J\5502 @b\5512 @z\5522 @\567-2 >>ONH2 -2 >\38- 2 >T\4.4222 >\-52.4512 >\-54.211-2 >\102.2142 >e\1.62562 >\3.52 "- --- \x:a'5%S 0< %"Systemn- "--   h}-$*}hh}($*58o;8;5o(858n:5858n:n:n:5 Arial--2 B,}h-1802 B|}h-90 2 B}h02 B}h902 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