Crystallization 2 and 3:
Lectin I (L-1) was concentrated to 10 mg/ml in 0.2 M glycine/HCl
buffer pH 2.5 for setups. To obtain L-I crystals in complex with
lactose and galactose, the sugar compound was added to the above
mentioned buffer to a concentration of 0.2 M. Crystals of L-I
complexed with lactose and galactose were obtained by hanging drop
vapour diffusion. Droplets of 1 μl protein and 1 μl reservoir solutions
were equilibrated against a reservoir of 0.75 ml of the reservoir
solution (30-40% saturated ammonium sulfate in 0.1 M glycine/HCl
buffer and 0.1 M lactose pH 2.5) to which was added in added 40 μl
dioxane. The crystallization conditions for the L-I galactose complex
differed only in the addition of 0.2 M galactose rather than lactose
to the protein solution.
Cryo-conditions for crystals 2 and 3:
The cryoprotectant solution contained 30-35% (v/v)
glycerol, 40% saturated ammonium sulphate, 0.1 M lactose/galactose
in 0.1 M glycine/HCl buffer at pH 2.5.
Example 2: lectin in complex with lactose
_exptl_crystal.crystal_id 2
_exptl_crystal.size_max ?
_exptl_crystal.size_mid ?
_exptl_crystal.size_min ?
_exptl_crystal_grow.crystal_id 2
_exptl_crystal_grow.method 'hanging-drop vapour diffusion'
_exptl_crystal_grow.temp_details 'ambient'
loop_
_pdbx_exptl_crystal_grow_sol.crystal_id
_pdbx_exptl_crystal_grow_sol.sol_id
_pdbx_exptl_crystal_grow_sol.volume
_pdbx_exptl_crystal_grow_sol.volume_units
_pdbx_exptl_crystal_grow_sol.pH
2 'protein' 1. 'microlitre' 2.5
2 'precipitant' 1. 'microlitre' 2.5
2 'reservoir' .79 'millilitre' 2.5
loop_
_pdbx_exptl_crystal_grow_comp.crystal_id
_pdbx_exptl_crystal_grow_comp.sol_id
_pdbx_exptl_crystal_grow_comp.comp_id
_pdbx_exptl_crystal_grow_comp.comp_name
_pdbx_exptl_crystal_grow_comp.conc
_pdbx_exptl_crystal_grow_comp.conc_range
_pdbx_exptl_crystal_grow_comp.conc_units
2 'protein' 1 'protein' 10 . 'milligrams_per_millilitre'
2 'protein' 2 'glycine/HCl' 0.1 . 'molar'
2 'protein' 3 'lactose/galactose' 0.2 . 'molar'
2 'reservoir' 1 'ammonium sulfate' . '30-40' 'percent_weight_by_volume'
2 'reservoir' 2 'glycine/HCl' 0.1 . 'molar'
2 'reservoir' 3 'lactose' 0.1 . 'molar'
2 'reservoir' 4 'dioxane' 5.0 . 'percent_volume_by_volume'
2 'precipitant' 1 'ammonium sulfate' . '30-40' 'percent_weight_by_volume'
2 'precipitant' 2 'glycine/HCl' 0.1 . 'molar'
2 'precipitant' 3 'lactose' 0.1 . 'molar'
2 'precipitant' 4 'dioxane' 5.0 . 'percent_volume_by_volume'
_pdbx_exptl_crystal_cryo_treatment.crystal_id 2
_pdbx_exptl_crystal_cryo_treatment.final_solution_details
; The cryoprotectant solution contained 30-35% (v/v) glycerol,
40% saturated ammonium sulfate, 0.1M lactose/galactose in 0.1M
glycine/HCl buffer at pH 2.5.
;
How this example will appear in the journal
| Crystallization |
| Crystallization method
|
Hanging-drop vapour diffusion
|
| Temperature
|
Ambient
|
| Crystallization solutions |
| Protein (1 μl)
|
Protein 10 mg ml-1,
glycine/HCl 0.1 M,
lactose/galactose 0.2 M, pH 2.5
|
| Precipitant (1 μl)
|
ammonium sulfate 30-40% (w/v),
glycine/HCl 0.1 M,
lactose 0.1 M,
dioxane 5.0% (w/v), pH 2.5
|
| Reservoir (0.79 ml)
|
ammonium sulfate 30-40% (w/v),
glycine/HCl 0.1 M,
lactose 0.1 M,
dioxane 5.0% (w/v), pH 2.5
|
| Cryo treatment
|
| Final cryoprotection solution
|
The cryoprotectant solution contained 30-35% (v/v)
glycerol, 40% saturated ammonium sulfate, 0.1M
lactose/galactose in 0.1M glycine/HCl buffer at pH 2.5
|
|
Example 3: lectin in complex with galactose
_exptl_crystal.crystal_id 3
_exptl_crystal.size_max ?
_exptl_crystal.size_mid ?
_exptl_crystal.size_min ?
_exptl_crystal_grow.crystal_id 3
_exptl_crystal_grow.method 'hanging-drop vapour diffusion'
_exptl_crystal_grow.temp_details 'ambient'
loop_
_pdbx_exptl_crystal_grow_sol.crystal_id
_pdbx_exptl_crystal_grow_sol.sol_id
_pdbx_exptl_crystal_grow_sol.volume
_pdbx_exptl_crystal_grow_sol.volume_units
_pdbx_exptl_crystal_grow_sol.pH
3 'protein' 1. 'microlitre' 2.5
3 'precipitant' 1. 'microlitre' 2.5
3 'reservoir' .79 'millilitre ' 2.5
loop_
_pdbx_exptl_crystal_grow_comp.crystal_id
_pdbx_exptl_crystal_grow_comp.sol_id
_pdbx_exptl_crystal_grow_comp.comp_id
_pdbx_exptl_crystal_grow_comp.comp_name
_pdbx_exptl_crystal_grow_comp.conc
_pdbx_exptl_crystal_grow_comp.conc_range
_pdbx_exptl_crystal_grow_comp.conc_units
3 'protein' 1 'protein' 10 . 'milligrams_per_millilitre'
3 'protein' 2 'glycine/HCl' 0.1 . 'molar'
3 'protein' 3 'lactose/galactose' 0.2 . 'molar'
3 'reservoir' 1 'ammonium sulfate' . '30-40' 'percent_weight_by_volume'
3 'reservoir' 2 'glycine/HCl' 0.1 . 'molar'
3 'reservoir' 3 'galactose' 0.1 . 'molar'
3 'reservoir' 4 'dioxane' 5.0 . 'percent_volume_by_volume'
3 'precipitant' 1 'ammonium sulfate' . '30-40' 'percent_weight_by_volume'
3 'precipitant' 2 'glycine/HCl' 0.1 . 'molar'
3 'precipitant' 3 'galactose' 0.1 . 'molar'
3 'precipitant' 4 'dioxane' 5.0 . 'percent_volume_by_volume'
_pdbx_exptl_crystal_cryo_treatment.crystal_id 3
_pdbx_exptl_crystal_cryo_treatment.final_solution_details
; The cryoprotectant solution contained 30-35% (v/v) glycerol,
40% saturated ammonium sulfate, 0.1M lactose/galactose in 0.1M
glycine/HCl buffer at pH 2.5.
;
How this example will appear in the journal
| Crystallization |
| Crystallization method
|
Hanging-drop vapour diffusion
|
| Temperature
|
Ambient
|
| Crystallization solutions |
| Protein (1 μl)
|
Protein 10 mg ml-1,
glycine/HCl 0.1 M,
lactose/galactose 0.2 M, pH 2.5
|
| Precipitant (1 μl)
|
ammonium sulfate 30-40% (w/v),
glycine/HCl 0.1 M,
lactose 0.1 M,
dioxane 5.0% (w/v), pH 2.5
|
| Reservoir (0.79 ml)
|
ammonium sulfate 30-40% (w/v),
glycine/HCl 0.1 M,
galactose 0.1 M,
dioxane 5.0% (w/v), pH 2.5
|
| Cryo treatment
|
| Final cryoprotection solution
|
The cryoprotectant solution contained 30-35% (v/v)
glycerol, 40% saturated ammonium sulfate, 0.1M
lactose/galactose in 0.1M glycine/HCl buffer at pH 2.5
|
|
|
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