Structure-based mutation analysis (PKU)

Finding the right structure

As proposed we searched the UNIProt entry for our protein and then selected the entry with the highest resolution and the lowest r-Value. In our case this is 1J8U which is the protein in a complex with its cosubstrate BH4. IN the following we will only use this structure, but we also list the results we found. <figtable id="uniprotresult">

Table with results from UNIProt with r-Value inserted

Entry	Method	Resolution (Å)	r-Value	Chain	Positions	PDBsum
1DMW	X-ray	2.00	0.200	A	118-424	[»]
1J8T	X-ray	1.70	0.197	A	103-427	[»]
1J8U	X-ray	1.50	0.157	A	103-427	[»]
1KW0	X-ray	2.50	0.220	A	103-427	[»]
1LRM	X-ray	2.10	0.211	A	103-427	[»]
1MMK	X-ray	2.00	0.199	A	103-427	[»]
1MMT	X-ray	2.00	0.213	A	103-427	[»]
1PAH	X-ray	2.00	0.176	A	117-424	[»]
1TDW	X-ray	2.10	0.206	A	117-424	[»]
1TG2	X-ray	2.20	0.213	A	117-424	[»]
2PAH	X-ray	3.10	0.251	A/B	118-452	[»]
3PAH	X-ray	2.00	0.175	A	117-424	[»]
4ANP	X-ray	2.11	0.204	A	104-427	[»]
4PAH	X-ray	2.00	0.169	A	117-424	[»]
5PAH	X-ray	2.10	0.163	A	117-424	[»]
6PAH	X-ray	2.15	0.171	A	117-424	[»]

</figtable> In <xr id="uniprotresult"/> there are all results according to which we selected 1J8U to be our reference for this weeks task. The corresponding line is marked in yellow.

1J8U

In order to know the structure of the protein and its important residues, we have a look at its structure with PyMol and visualize the BH4 and the Fe-ion with the most important residues.

<figure id="1J8Uwhole">

Rendering of the overall structures of 1J8U using PyMol. The protein is colored cyan overall, whereas the Fe-atom is colored red and the important residues are shown as sticks and colored in the element-based fashion. Binding is shown with yellow strokes, if the distance is bigger than 1.5 Å

</figure>

<figure id="1J8Uclose">

Rendering of a close-up of the structures of 1J8U using PyMol. The protein is colored cyan overall, whereas the Fe-atom is colored red and the important residues are shown as sticks and colored in the element-based fashion. Binding is shown with yellow strokes, if the distance is bigger than 1.5 Å

</figure>

Mutations

As you probably know from last weeks dataset the first two mutations are located before residue 103 and therefore not contained in the structure. We will change them according to the premise, that the number of diseasecausing and non-diseasecausing mutations are equal.
We propose the following dataset, chosen mostly from well known SNPs from OMIM. They include mutations causing no reported effect, the mild related hyperphenylalaninemia (reduced activity, but functional enzyme) and phenylketonuria.

SNP	effect	prediction	validation
ARG158GLN	disease causing
GLN172HIS	non-disease
ARG243GLN	disease causing
LEU255SER	disease causing
MET276VAL	non-disease
ARG297CYS	disease causing
ALA322GLY	hyperphenylalaninemia
GLU330ASP	disease causing			GLY337VAL	disease causing
ARG408TRP	disease causing