File size: 13,291 Bytes
d766458
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
{
  "cells": [
    {
      "cell_type": "markdown",
      "metadata": {
        "id": "view-in-github",
        "colab_type": "text"
      },
      "source": [
        "<a href=\"https://colab.research.google.com/github/sokrypton/ColabDesign/blob/v1.1.1/af/design.ipynb\" target=\"_parent\"><img src=\"https://colab.research.google.com/assets/colab-badge.svg\" alt=\"Open In Colab\"/></a>"
      ]
    },
    {
      "cell_type": "markdown",
      "metadata": {
        "id": "OA2k3sAYuiXe"
      },
      "source": [
        "#AfDesign (v1.1.1)\n",
        "Backprop through AlphaFold for protein design.\n",
        "\n",
        "**WARNING**\n",
        "1.   This notebook is in active development and was designed for demonstration purposes only.\n",
        "2.   Using AfDesign as the only \"loss\" function for design might be a bad idea, you may find adversarial sequences (aka. sequences that trick AlphaFold)."
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "cellView": "form",
        "id": "-AXy0s_4cKaK"
      },
      "outputs": [],
      "source": [
        "#@title setup\n",
        "%%time\n",
        "import os\n",
        "if not os.path.isdir(\"params\"):\n",
        "  # get code\n",
        "  os.system(\"pip -q install git+https://github.com/sokrypton/ColabDesign.git@v1.1.1\")\n",
        "  # for debugging\n",
        "  os.system(\"ln -s /usr/local/lib/python3.*/dist-packages/colabdesign colabdesign\")\n",
        "  # download params\n",
        "  os.system(\"mkdir params\")\n",
        "  os.system(\"apt-get install aria2 -qq\")\n",
        "  os.system(\"aria2c -q -x 16 https://storage.googleapis.com/alphafold/alphafold_params_2022-12-06.tar\")\n",
        "  os.system(\"tar -xf alphafold_params_2022-12-06.tar -C params\")\n",
        "\n",
        "import warnings\n",
        "warnings.simplefilter(action='ignore', category=FutureWarning)\n",
        "\n",
        "import os\n",
        "from colabdesign import mk_afdesign_model, clear_mem\n",
        "from IPython.display import HTML\n",
        "from google.colab import files\n",
        "import numpy as np\n",
        "\n",
        "def get_pdb(pdb_code=\"\"):\n",
        "  if pdb_code is None or pdb_code == \"\":\n",
        "    upload_dict = files.upload()\n",
        "    pdb_string = upload_dict[list(upload_dict.keys())[0]]\n",
        "    with open(\"tmp.pdb\",\"wb\") as out: out.write(pdb_string)\n",
        "    return \"tmp.pdb\"\n",
        "  elif os.path.isfile(pdb_code):\n",
        "    return pdb_code\n",
        "  elif len(pdb_code) == 4:\n",
        "    os.system(f\"wget -qnc https://files.rcsb.org/view/{pdb_code}.pdb\")\n",
        "    return f\"{pdb_code}.pdb\"\n",
        "  else:\n",
        "    os.system(f\"wget -qnc https://alphafold.ebi.ac.uk/files/AF-{pdb_code}-F1-model_v3.pdb\")\n",
        "    return f\"AF-{pdb_code}-F1-model_v3.pdb\""
      ]
    },
    {
      "cell_type": "markdown",
      "metadata": {
        "id": "UUfKrOzT0gOS"
      },
      "source": [
        "# fixed backbone design (fixbb)\n",
        "For a given protein backbone, generate/design a new sequence that AlphaFold thinks folds into that conformation. "
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "qLd1DsnKzxBJ"
      },
      "outputs": [],
      "source": [
        "clear_mem()\n",
        "af_model = mk_afdesign_model(protocol=\"fixbb\")\n",
        "af_model.prep_inputs(pdb_filename=get_pdb(\"1TEN\"), chain=\"A\")\n",
        "\n",
        "print(\"length\",  af_model._len)\n",
        "print(\"weights\", af_model.opt[\"weights\"])"
      ]
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.restart()\n",
        "af_model.design_3stage()"
      ],
      "metadata": {
        "id": "u0AwskJ84NGx"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.plot_traj()  "
      ],
      "metadata": {
        "id": "8FB1v7dn1LL6"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "YEApO8YzBoS0"
      },
      "outputs": [],
      "source": [
        "af_model.save_pdb(f\"{af_model.protocol}.pdb\")\n",
        "af_model.plot_pdb()"
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "cW1KQiHKJpfp"
      },
      "outputs": [],
      "source": [
        "HTML(af_model.animate())"
      ]
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.get_seqs()"
      ],
      "metadata": {
        "id": "YDrChASGVUUx"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "markdown",
      "source": [
        "# hallucination\n",
        "For a given length, generate/hallucinate a protein sequence that AlphaFold thinks folds into a well structured protein (high plddt, low pae, many contacts)."
      ],
      "metadata": {
        "id": "qLwS2s_xcjRI"
      }
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "sZnYfCbfEvol"
      },
      "outputs": [],
      "source": [
        "clear_mem()\n",
        "af_model = mk_afdesign_model(protocol=\"hallucination\")\n",
        "af_model.prep_inputs(length=100)\n",
        "\n",
        "print(\"length\",af_model._len)\n",
        "print(\"weights\",af_model.opt[\"weights\"])"
      ]
    },
    {
      "cell_type": "code",
      "source": [
        "# pre-design with gumbel initialization and softmax activation\n",
        "af_model.restart(mode=\"gumbel\")\n",
        "af_model.design_soft(50)\n",
        "\n",
        "# three stage design  \n",
        "af_model.set_seq(af_model.aux[\"seq\"][\"pseudo\"])\n",
        "af_model.design_3stage(50,50,10)"
      ],
      "metadata": {
        "id": "f76xqCkw0vj9"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "A1GxeLZdTTya"
      },
      "outputs": [],
      "source": [
        "af_model.save_pdb(f\"{af_model.protocol}.pdb\")\n",
        "af_model.plot_pdb()"
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "L2E9Tn2Acchj"
      },
      "outputs": [],
      "source": [
        "HTML(af_model.animate())"
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "YSKWYu0_GlUH"
      },
      "outputs": [],
      "source": [
        "af_model.get_seqs()"
      ]
    },
    {
      "cell_type": "markdown",
      "metadata": {
        "id": "dXfm4B8ISLuL"
      },
      "source": [
        "# binder hallucination\n",
        "For a given protein target and protein binder length, generate/hallucinate a protein binder sequence AlphaFold thinks will bind to the target structure.\n",
        "To do this, we minimize PAE and maximize number of contacts at the interface and within the binder, and we maximize pLDDT of the binder.\n",
        "By default, AlphaFold-ptm with residue index offset hack is used. To enable AlphaFold-multimer set: mk_afdesign_model(use_multimer=True).\n"
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "3XLJPiRKx5Mw"
      },
      "outputs": [],
      "source": [
        "clear_mem()\n",
        "af_model = mk_afdesign_model(protocol=\"binder\")\n",
        "af_model.prep_inputs(pdb_filename=get_pdb(\"4MZK\"), chain=\"A\", binder_len=19)\n",
        "\n",
        "print(\"target_length\",af_model._target_len)\n",
        "print(\"binder_length\",af_model._binder_len)\n",
        "print(\"weights\",af_model.opt[\"weights\"])"
      ]
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "u6VxjuinyCZa"
      },
      "outputs": [],
      "source": [
        "af_model.restart()\n",
        "af_model.design_3stage(100,100,10)"
      ]
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.save_pdb(f\"{af_model.protocol}.pdb\")\n",
        "af_model.plot_pdb()"
      ],
      "metadata": {
        "id": "sTlS7_L8Zfwf"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "execution_count": null,
      "metadata": {
        "id": "9cARoviGyIKb"
      },
      "outputs": [],
      "source": [
        "HTML(af_model.animate())"
      ]
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.get_seqs()"
      ],
      "metadata": {
        "id": "RzE137NDZdZc"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "markdown",
      "source": [
        "#**ADVANCED**"
      ],
      "metadata": {
        "id": "SK0IJLoen_EC"
      }
    },
    {
      "cell_type": "markdown",
      "source": [
        "## partial hallucination + custom Radius of Gyration (rg) loss\n",
        "mix supervised (fixbb) and unsupervised (hallucination) losses to constrain the halluciation process."
      ],
      "metadata": {
        "id": "zl6JGTUzXRnk"
      }
    },
    {
      "cell_type": "code",
      "source": [
        "import jax\n",
        "import jax.numpy as jnp\n",
        "from colabdesign.af.alphafold.common import residue_constants\n",
        "\n",
        "# first off, let's implement a custom Radius of Gyration loss function\n",
        "def rg_loss(inputs, outputs):\n",
        "  positions = outputs[\"structure_module\"][\"final_atom_positions\"]\n",
        "  ca = positions[:,residue_constants.atom_order[\"CA\"]]\n",
        "  center = ca.mean(0)\n",
        "  rg = jnp.sqrt(jnp.square(ca - center).sum(-1).mean() + 1e-8)\n",
        "  rg_th = 2.38 * ca.shape[0] ** 0.365\n",
        "  rg = jax.nn.elu(rg - rg_th)\n",
        "  return {\"rg\":rg}"
      ],
      "metadata": {
        "id": "spec3m8BlGer"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "clear_mem()\n",
        "af_model = mk_afdesign_model(protocol=\"partial\",\n",
        "                             loss_callback=rg_loss, # add rg_loss\n",
        "                             use_templates=False)   # set True to constrain positions using template input\n",
        "\n",
        "af_model.opt[\"weights\"][\"rg\"] = 0.1  # optional: specify weight for rg_loss\n",
        "\n",
        "af_model.prep_inputs(pdb_filename=get_pdb(\"6MRR\"),\n",
        "                     chain=\"A\",\n",
        "                     pos=\"3-30,33-68\",  # define positions to contrain\n",
        "                     length=100)          # total length if different from input pdb\n",
        "\n",
        "af_model.rewire(loops=[36]) # set loop length between segments                     "
      ],
      "metadata": {
        "id": "h_BvzwbAKo6V"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "# initialize with wildtype seq, fill in the rest with soft_gumbel distribution\n",
        "af_model.restart(mode=[\"soft\",\"gumbel\",\"wildtype\"])\n",
        "af_model.design_3stage(100, 100, 10)"
      ],
      "metadata": {
        "id": "5Unr9u2GYKRD"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.save_pdb(f\"{af_model.protocol}.pdb\")\n",
        "af_model.plot_pdb()"
      ],
      "metadata": {
        "id": "BFweaqNWYuF0"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "HTML(af_model.animate())"
      ],
      "metadata": {
        "id": "GSu2lB9HYw-t"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [
        "af_model.get_seqs()"
      ],
      "metadata": {
        "id": "2EG2t2_KY4Td"
      },
      "execution_count": null,
      "outputs": []
    },
    {
      "cell_type": "code",
      "source": [],
      "metadata": {
        "id": "rTGKbhsI0t8k"
      },
      "execution_count": null,
      "outputs": []
    }
  ],
  "metadata": {
    "accelerator": "GPU",
    "colab": {
      "collapsed_sections": [
        "q4qiU9I0QHSz"
      ],
      "name": "design.ipynb",
      "provenance": [],
      "include_colab_link": true
    },
    "kernelspec": {
      "display_name": "Python 3",
      "name": "python3"
    },
    "language_info": {
      "name": "python"
    }
  },
  "nbformat": 4,
  "nbformat_minor": 0
}