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+# Instruction of running structural predictions with cross-link data via [AlphaLink2](https://github.com/Rappsilber-Laboratory/AlphaLink2/tree/main)
+## Introduction
+As [Stahl et al., 2023](https://www.nature.com/articles/s41587-023-01704-z) showed, integrating cross-link data with AlphaFold could improve the modelling quality in 
+some challenging cases. Thus AlphaPulldown has integrated [AlphaLink2](https://github.com/Rappsilber-Laboratory/AlphaLink2/tree/main) pipeline 
+and allows the user to combine cross-link data with AlphaFold Multimer inference, without the need of calculating MSAs from the scratch again.
+
+In addition, this integration retains all the other benefits from AlphaPulldown, such as the interface for fragmenting protein into regions; automatically 
+generating PAE plots after the predictions etc.
+
+## 1st step: configure the Conda environment
+After you initialise the same conda environment, where you normally run AlphaPulldown, firstly, you need to compile [UniCore](https://github.com/dptech-corp/Uni-Core).
+
+```bash
+git clone https://github.com/dptech-corp/Uni-Core.git
+cd Uni-Core
+python setup.py install --disable-cuda-ext
+
+# test whether unicore is successfully installed
+python -c "import unicore"
+```
+You may see the following warning but it's fine: 
+```
+fused_multi_tensor is not installed corrected
+fused_rounding is not installed corrected
+fused_layer_norm is not installed corrected
+fused_softmax is not installed corrected
+```
+Next, make sure you have PyTorch corresponding to the CUDA version installed. For example, [PyTorch 1.13.0+cu117](https://pytorch.org/get-started/previous-versions/) 
+and CUDA/11.7.0 
+## 2nd step: download AlphaLink2 checkpoint 
+Now please download the PyTorch checkpoints from [Zenodo](https://zenodo.org/records/8007238), unzip it, then you should obtain a file named: ```AlphaLink-Multimer_SDA_v3.pt```
+
+## 3rd step: prepare cross-link input data
+As instructed by [AlphaLink2](https://github.com/Rappsilber-Laboratory/AlphaLink2/tree/main), information of cross-linked residues
+between 2 proteins, inter-protein crosslinks A->B 1,50 and 30,80 and an FDR=20%, should look like: 
+```
+{'protein_A': {'protein_B': [(1, 50, 0.2), (30, 80, 0.2)]}}
+```
+and intra-protein crosslinks follow the same format: 
+```
+{'protein_A': {'protein_A': [(5, 20, 0.2)]}}
+```
+The keys in these dictionaries should be the same as your pickle files created by [the first stage of AlphaPulldown](https://github.com/KosinskiLab/AlphaPulldown/blob/main/example_1.md). e.g. you should have ```protein_A.pkl``` 
+and ```protein_B.pkl``` already calculated. 
+
+Dictionaries like these should be stored in **```.pkl.gz```** files and provided to AlphaPulldown in the next step. You can use the script from [AlphaLink2](https://github.com/Rappsilber-Laboratory/AlphaLink2/tree/main)
+to prepare these pickle files. 
+### **NB** The dictionaries are 0-indexed, i.e., residues start from 0.
+
+## 4th step: run with AlphaLink2 prediction via AlphaPulldown
+Within the same conda environment, run in e.g. ```custom``` mode:
+```bash
+run_multimer_jobs.py --mode=custom \
+--num_predictions_per_model=1 \
+--output_path=/scratch/scratch/user/output/models \
+--data_dir=/g/alphafold/AlphaFold_DBs/2.3.0/ \
+--protein_lists=custom.txt \
+--monomer_objects_dir=/scratch/user/output/features \
+--job_index=$SLURM_ARRAY_TASK_ID --alphalink_weight=/scratch/user/alphalink_weights/AlphaLink-Multimer_SDA_v3.pt \
+--use_alphalink=True --crosslinks=/path/to/crosslinks.pkl.gz 
+```
+The other modes provided by AlphaPulldown also work in the same way.
+
+
+