IGDRPT (Indian Genetic Disease Risk Prediction Tool)

Disease Name: Beta Thalassemia

1 .Inheritance

Autosomal Recessive

2. Gene Name

HBB

3. Protein Name

Hemoglobin subunit beta

4. Chromosomal Location

11q15.5

5. Protein Sequence

>sp|P68871|HBB_HUMAN Hemoglobin subunit beta OS=Homo sapiens OX=9606 GN=HBB PE=1 SV=2

MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPK

VKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFG

KEFTPPVQAAYQKVVAGVANALAHKYH

6. Gene Sequence

>NM_000518.4 Homo sapiens hemoglobin subunit beta (HBB), mRNA

ACATTTGCTTCTGACACAACTGTGTTCACTAGCAACCTCAAACAGACACCATGGTGCATCTGACTCCTGA

GGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAGTTGGTGGTGAGGCCCTGGGC

AGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGGGGATCTGTCCACTCCTGATG

CTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGTGCCTTTAGTGATGGCCTGGC

TCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACTGTGACAAGCTGCACGTGGAT

CCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCATCACTTTGGCAAAGAATTCA

CCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAATGCCCTGGCCCACAAGTATCA

CTAAGCTCGCTTTCTTGCTGTCCAATTTCTATTAAAGGTTCCTTTGTTCCCTAAGTCCAACTACTAAACT

GGGGGATATTATGAAGGGCCTTGAGCATCTGGATTCTGCCTAATAAAAAACATTTATTTTCATTGC

7. Motif

Pfam (1 motif)

Pfam	Position(Independent E-value)	Description
Globin	27..142(4.6e-33)	PF00042, Globin

8. Evolutionary Relationship

Evolutionary analysis by Maximum Likelihood method

The evolutionary history was inferred by using the Maximum Likelihood method and JTT matrix-based model [1]. The tree with the highest log likelihood (-1343.01) is shown. Initial tree(s) for the heuristic search were obtained automatically by applying Neighbor-Join and BioNJ algorithms to a matrix of pairwise distances estimated using a JTT model, and then selecting the topology with superior log likelihood value. The proportion of sites where at least 1 unambiguous base is present in at least 1 sequence for each descendent clade is shown next to each internal node in the tree. This analysis involved 100 amino acid sequences. There were a total of 148 positions in the final dataset.

9. SNP

SNP	COORDINATE	AMINO ACID CHANGE	GENE ID	TRANSCRIPT ID	PROTEIN ID	SIFT SCORE	SIFT MEDIAN	SIFT PREDICTION
rs334	5248232	E7V	ENSG00000244734	ENST00000335295	ENSP00000333994	0.063	2.49	TOLERATED
rs334	5248232	E7G	ENSG00000244734	ENST00000335295	ENSP00000333994	0.117	2.49	TOLERATED
rs334	5248232	E7A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.58	2.49	TOLERATED
rs1135071	5248029	R31S	ENSG00000244734	ENST00000335295	ENSP00000333994	0.029	2.49	DELETERIOUS
rs1135071	5248029	R31S	ENSG00000244734	ENST00000335295	ENSP00000333994	0.029	2.49	DELETERIOUS
rs1141387	5248019	V35F	ENSG00000244734	ENST00000335295	ENSP00000333994	0.02	2.49	DELETERIOUS
rs1141387	5248019	V35L	ENSG00000244734	ENST00000335295	ENSP00000333994	0.003	2.49	DELETERIOUS
rs11549407	5248004	Q40*	ENSG00000244734	ENST00000335295	ENSP00000333994
rs11549407	5248004	Q40E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.49	DELETERIOUS
rs11549407	5248004	Q40K	ENSG00000244734	ENST00000335295	ENSP00000333994	0.396	2.49	TOLERATED
rs33910209	5246878	Q132E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.277	2.51	TOLERATED
rs33910209	5246878	Q132K	ENSG00000244734	ENST00000335295	ENSP00000333994	0.02	2.51	DELETERIOUS
rs33910569	5246889	Q128R	ENSG00000244734	ENST00000335295	ENSP00000333994	0.018	2.51	DELETERIOUS
rs33910569	5246889	Q128P	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.51	DELETERIOUS
rs33913712	5247851	E91*	ENSG00000244734	ENST00000335295	ENSP00000333994
rs33913712	5247851	E91K	ENSG00000244734	ENST00000335295	ENSP00000333994	0.258	2.5	TOLERATED
rs33914944	5247807	R105S	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.5	DELETERIOUS
rs33914944	5247807	R105S	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.5	DELETERIOUS
rs33915112	5248172	E27V	ENSG00000244734	ENST00000335295	ENSP00000333994	0.016	2.49	DELETERIOUS
rs33915112	5248172	E27G	ENSG00000244734	ENST00000335295	ENSP00000333994	0.013	2.49	DELETERIOUS
rs33915112	5248172	E27A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.104	2.49	TOLERATED
rs33916412	5248166	L29R	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.49	DELETERIOUS
rs33916412	5248166	L29P	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.49	DELETERIOUS
rs33916412	5248166	L29Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.49	DELETERIOUS
rs33917628	5247853	S90T	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33917628	5247853	S90N	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33917785	5247847	L92R	ENSG00000244734	ENST00000335295	ENSP00000333994	0.03	2.5	DELETERIOUS
rs33917785	5247847	L92P	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.5	DELETERIOUS
rs33918338	5246841	H144L	ENSG00000244734	ENST00000335295	ENSP00000333994	0.12	2.5	TOLERATED
rs33918338	5246841	H144R	ENSG00000244734	ENST00000335295	ENSP00000333994	1	2.5	TOLERATED
rs33918338	5246841	H144P	ENSG00000244734	ENST00000335295	ENSP00000333994	0.068	2.5	TOLERATED
rs33918343	5247919	V68G	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33918343	5247919	V68A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.012	2.5	DELETERIOUS
rs33918343	5247919	V68E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.017	2.5	DELETERIOUS
rs33921821	5246844	A143V	ENSG00000244734	ENST00000335295	ENSP00000333994	0.022	2.5	DELETERIOUS
rs33921821	5246844	A143D	ENSG00000244734	ENST00000335295	ENSP00000333994	0.003	2.5	DELETERIOUS
rs33922842	5247992	E44*	ENSG00000244734	ENST00000335295	ENSP00000333994
rs33922842	5247992	E44Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0.224	2.5	TOLERATED
rs33922842	5247992	E44K	ENSG00000244734	ENST00000335295	ENSP00000333994	0.397	2.5	TOLERATED
rs33922873	5247932	H64Y	ENSG00000244734	ENST00000335295	ENSP00000333994	0.005	2.51	DELETERIOUS
rs33922873	5247932	H64N	ENSG00000244734	ENST00000335295	ENSP00000333994	0.007	2.51	DELETERIOUS
rs33924146	5247995	F43V	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.49	DELETERIOUS
rs33924146	5247995	F43L	ENSG00000244734	ENST00000335295	ENSP00000333994	0.004	2.49	DELETERIOUS
rs33924775	5247845	H93Y	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33924775	5247845	H93D	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33924775	5247845	H93N	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33925391	5246892	V127G	ENSG00000244734	ENST00000335295	ENSP00000333994	0.014	2.51	DELETERIOUS
rs33925391	5246892	V127A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.329	2.51	TOLERATED
rs33925391	5246892	V127E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.011	2.51	DELETERIOUS
rs33926764	5248227	K9E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.002	2.49	DELETERIOUS
rs33926764	5248227	K9Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0.003	2.49	DELETERIOUS
rs33929459	5248182	V24F	ENSG00000244734	ENST00000335295	ENSP00000333994	0.103	2.48	TOLERATED
rs33929459	5248182	V24I	ENSG00000244734	ENST00000335295	ENSP00000333994	0.981	2.48	TOLERATED
rs33930165	5248233	E7Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0.182	2.49	TOLERATED
rs33930165	5248233	E7K	ENSG00000244734	ENST00000335295	ENSP00000333994	0.408	2.49	TOLERATED
rs33930702	5248249	M1I	ENSG00000244734	ENST00000335295	ENSP00000333994	0.02	2.77	DELETERIOUS
rs33930702	5248249	M1I	ENSG00000244734	ENST00000335295	ENSP00000333994	0.02	2.77	DELETERIOUS
rs33930702	5248249	M1I	ENSG00000244734	ENST00000335295	ENSP00000333994	0.02	2.77	DELETERIOUS
rs33930977	5246933	C113W	ENSG00000244734	ENST00000335295	ENSP00000333994	0.001	2.5	DELETERIOUS
rs33930977	5246933	C113*	ENSG00000244734	ENST00000335295	ENSP00000333994
rs33931779	5247940	V61A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.006	2.51	DELETERIOUS
rs33931779	5247940	V61E	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.51	DELETERIOUS
rs33933298	5247827	V99L	ENSG00000244734	ENST00000335295	ENSP00000333994	0.007	2.5	DELETERIOUS
rs33933298	5247827	V99M	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33937393	5247817	E102G	ENSG00000244734	ENST00000335295	ENSP00000333994	0.154	2.5	TOLERATED
rs33937393	5247817	E102A	ENSG00000244734	ENST00000335295	ENSP00000333994	0.352	2.5	TOLERATED
rs33940051	5247875	K83E	ENSG00000244734	ENST00000335295	ENSP00000333994	0.019	2.51	DELETERIOUS
rs33940051	5247875	K83Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0.017	2.51	DELETERIOUS
rs33940204	5247856	L89R	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33940204	5247856	L89P	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33941844	5246952	L107R	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33941844	5246952	L107P	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33941844	5246952	L107Q	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.5	DELETERIOUS
rs33941849	5248250	M1R	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.77	DELETERIOUS
rs33941849	5248250	M1T	ENSG00000244734	ENST00000335295	ENSP00000333994	0.001	2.77	DELETERIOUS
rs33941849	5248250	M1K	ENSG00000244734	ENST00000335295	ENSP00000333994	0	2.77	DELETERIOUS

Evolutionary analysis of coding SNPs

SNP	Substitution	Probability of deleterious effect	Prediction Score
rs334	E7V	probably damaging	0.57
rs334	E7G	probably damaging	0.57
rs334	E7A	probably damaging	0.57
rs1135071	R31S	probably damaging	0.85
rs1135071	R31S	probably damaging	0.85
rs1141387	V35F	probably damaging	0.57
rs1141387	V35L	probably damaging	0.57
rs11549407	Q40E	probably damaging	0.57
rs11549407	Q40K	probably damaging	0.57
rs11549407	Q132E	possibly damaging	0.5
rs33910209	Q132K	possibly damaging	0.5
rs33910209	Q128R	probably damaging	0.57
rs33910569	Q128P	probably damaging	0.57
rs33910569	E91K	possibly damaging	0.5
rs33913712	R105S	possibly damaging	0.5
rs33913712	R105S	possibly damaging	0.5
rs33914944	E27V	probably damaging	0.57
rs33914944	E27G	probably damaging	0.57
rs33915112	E27A	probably damaging	0.57
rs33915112	L29R	probably damaging	0.74
rs33915112	L29P	probably damaging	0.74
rs33916412	L29Q	probably damaging	0.74
rs33916412	S90T	probably damaging	0.57
rs33916412	S90N	probably damaging	0.57
rs33917628	L92R	possibly damaging	0.5
rs33917628	L92P	possibly damaging	0.5
rs33917785	H144L	possibly damaging	0.5
rs33917785	H144R	possibly damaging	0.5
rs33918338	H144P	possibly damaging	0.5
rs33918338	V68G	probably damaging	0.74
rs33918338	V68A	probably damaging	0.74
rs33918343	V68E	probably damaging	0.74
rs33918343	A143V	possibly damaging	0.5
rs33918343	A143D	possibly damaging	0.5
rs33921821	E44Q	probably benign	0.19
rs33921821	E44K	probably benign	0.19
rs33922842	H64Y	probably damaging	0.85
rs33922842	H64N	probably damaging	0.85
rs33922842	F43V	probably damaging	0.74
rs33922873	F43L	probably damaging	0.74
rs33922873	H93Y	probably damaging	0.85
rs33924146	H93D	probably damaging	0.85
rs33924146	H93N	probably damaging	0.85
rs33924775	V127G	possibly damaging	0.5
rs33924775	V127A	possibly damaging	0.5
rs33924775	V127E	possibly damaging	0.5
rs33925391	K9E	possibly damaging	0.5
rs33925391	K9Q	possibly damaging	0.5
rs33925391	V24F	probably benign	0.19
rs33926764	V24I	probably benign	0.19
rs33926764	E7Q	probably damaging	0.57
rs33929459	E7K	probably damaging	0.57
rs33929459	M1I	probably damaging	0.74
rs33930165	M1I	probably damaging	0.74
rs33930165	M1I	probably damaging	0.74
rs33930702	C113W	probably benign	0.19
rs33930702	V61A	probably damaging	0.57
rs33930702	V61E	probably damaging	0.57
rs33930977	V99L	probably damaging	0.57
rs33930977	V99M	probably damaging	0.57
rs33931779	E102G	possibly damaging	0.5
rs33931779	E102A	possibly damaging	0.5
rs33933298	K83E	probably damaging	0.57
rs33933298	K83Q	probably damaging	0.57
rs33937393	L89R	probably damaging	0.57
rs33937393	L89P	probably damaging	0.57
rs33940051	L107R	probably damaging	0.57
rs33940051	L107P	probably damaging	0.57
rs33940204	L107Q	probably damaging	0.57
rs33940204	M1R	probably damaging	0.74
rs33941844	M1T	probably damaging	0.74
rs33941844	M1K	probably damaging	0.74

Provean

SNP Id’s	Amino Acid Change	probability	Prediction	Score
rs334	E7V	Deleterious	Damaging	-4.71
rs334	E7G	Deleterious	Tolerated	-3.69
rs334	E7A	Deleterious	Damaging	-4.23
rs1135071	R31S	Deleterious	Damaging	-4.23
rs1135071	R31S	Deleterious	Damaging	-4.27
rs1141387	V35F	Neutral	Damaging	-2.45
rs1141387	V35L	Deleterious	Damaging	-2.52
rs11549407	Q40E	Deleterious	Damaging	-3.43
rs11549407	Q40K	Neutral	Tolerated	-1.74
rs33910209	Q132E	Neutral	Tolerated	-1.74
rs33910209	Q132K	Deleterious	Damaging	-2.66
rs33910569	Q128R	Deleterious	Damaging	-3.99
rs33910569	Q128P	Deleterious	Damaging	-3.43
rs33913712	E91K	Deleterious	Damaging	-3.51
rs33914944	R105S	Deleterious	Damaging	-3.51
rs33914944	R105S	Deleterious	Damaging	-5.55
rs33915112	E27V	Deleterious	Damaging	-5.45
rs33915112	E27G	Deleterious	Damaging	-4.57
rs33915112	E27A	Deleterious	Damaging	-5.13
rs33916412	L29R	Deleterious	Damaging	-5.89
rs33916412	L29P	Deleterious	Damaging	-5.13
rs33916412	L29Q	Deleterious	Damaging	-2.87
rs33917628	S90T	Deleterious	Damaging	-2.86
rs33917628	S90N	Deleterious	Damaging	-5.37
rs33917785	L92R	Deleterious	Damaging	-6.33
rs33917785	L92P	Deleterious	Damaging	-5.09
rs33918338	H144L	Deleterious	Damaging	-5.09
rs33918338	H144R	Deleterious	Damaging	-4.35
rs33918338	H144P	Deleterious	Damaging	-6.39
rs33918343	V68G	Deleterious	Damaging	-3.62
rs33918343	V68A	Deleterious	Damaging	-5.34
rs33918343	V68E	Deleterious	Damaging	-2.81
rs33921821	A143V	Deleterious	Damaging	-3.86
rs33921821	A143D	Neutral	Damaging	-1.31
rs33922842	E44Q	Neutral	Damaging	-1.94
rs33922842	E44K	Deleterious	Damaging	-5.66
rs33922873	H64Y	Deleterious	Damaging	-6.61
rs33922873	H64N	Deleterious	Damaging	-6.4
rs33924146	F43V	Deleterious	Damaging	-5.48
rs33924146	F43L	Deleterious	Damaging	-5.65
rs33924775	H93Y	Deleterious	Damaging	-8.48
rs33924775	H93D	Deleterious	Damaging	-6.6
rs33924775	H93N	Deleterious	Damaging	-2.82
rs33925391	V127G	Neutral	Tolerated	-0.82
rs33925391	V127A	Deleterious	Damaging	-2.62
rs33925391	V127E	Deleterious	Damaging	-3.25
rs33926764	K9E	Deleterious	Damaging	-3.19
rs33926764	K9Q	Deleterious	Damaging	-2.73
rs33929459	V24F	Neutral	Tolerated	-0.24
rs33929459	V24I	Neutral	Damaging	-2.02
rs33930165	E7Q	Deleterious	Damaging	-2.57
rs33930165	E7K	Neutral	Damaging	-2.07
rs33930702	M1I	Neutral	Damaging	-2.07
rs33930702	M1I	Neutral	Damaging	-2.07

SNAP 2

Wildetype Amino Acid	Position	Variant Amino Acid	Predicted Effect	Score	Expected Accuracy
E	7	V	Effect	67	80%
E	7	G	Effect	87	75%
E	7	A	Effect	76	65%
R	31	S	Effect	98	25%
R	31	S	Neutral	65	45%
V	35	F	Neutral	64	76%
V	35	L	Effect	53	78%
Q	40	E	Effect	76	90%
Q	40	K	Neutral	98	80%
Q	132	E	Effect	56	46%
Q	132	K	Effect	45	54%
Q	128	R	Effect	45	43%
Q	128	P	Neutral	65	39%
E	91	K	Neutral	88	90%
R	105	S	Neutral	76	89%
R	105	S	Neutral	73	70%
E	27	V	Effect	56	80%
E	27	G	Effect	36	70%
E	27	A	Effect	66	35%
L	29	R	Neutral	77	78%
L	29	P	Neutral	67	35%
L	90	Q	Effect	-25	96%
S	90	T	Effect	4	25%
L	92	N	Effect	5	45%
L	92	R	Neutral	23	35%
H	144	P	Effect	7	45%
H	144	L	Effect	87	76%
H	144	R	Effect	67	78%
V	68	P	Effect	98	90%
V	68	G	Effect	90	80%
V	68	A	Effect	75	46%
A	143	E	Neutral	75	54%
A	143	V	Neutral	34	43%
E	44	D	Neutral	45	39%
E	44	Q	Neutral	87	90%
H	64	K	Effect	76	89%
H	64	Y	Effect	65	70%
F	43	N	Neutral	34	80%
F	43	V	Neutral	45	70%
H	93	L	Effect	76	35%
H	93	Y	Effect	56	98%

Polyphen 2

Rs_Id	Mutation Probability	Score
rs334	Probably damaging	0.98
rs334	Possibly damaging	0.98
rs334	Possibly damaging	1.00
rs1135071	Possibly damaging	0.98
rs1135071	Probably damaging	0.98
rs1141387	Possibly damaging	0.98
rs1141387	Possibly damaging	1.00
rs11549407	Possibly damaging	0.98
rs11549407	Possibly damaging	1.00
rs33910209	Possibly damaging	0.98
rs33910209	Probably damaging	0.999
rs33910569	Possibly damaging	0.98
rs33910569	Possibly damaging	1.00
rs33913712	Possibly damaging	1.00
rs33914944	Probably damaging	0.98
rs33914944	Possibly damaging	0.98
rs33915112	Possibly damaging	1.00
rs33915112	Probably damaging	0.98
rs33915112	Probably damaging	0.98
rs33916412	Possibly damaging	0.98
rs33916412	Possibly damaging	1.00
rs33916412	Possibly damaging	0.98
rs33917628	Possibly damaging	1.00
rs33917628	Possibly damaging	0.98
rs33917785	Probably damaging	0.98
rs33917785	Probably damaging	0.999
rs33918338	Probably damaging	0.999
rs33918338	Possibly damaging	1.00
rs33918338	Possibly damaging	1.00
rs33918343	Possibly damaging	0.98
rs33918343	Probably damaging	0.999
rs33918343	Possibly damaging	0.98
rs33921821	Possibly damaging	1.00
rs33921821	Probably damaging	0.98
rs33922842	Possibly damaging	0.98
rs33922842	Possibly damaging	1.00
rs33922873	Possibly damaging	1.00
rs33922873	Probably damaging	0.98

Hope Prediction

Rs_ids	Mutation	Mapping Issue	AA variant	Function. Impact	Score
rs334	HBB_E7V	Uniprot residue:H	E7V
rs334	HBB_E7G		E7G	High	9.77
rs334	HBB_E7A		E7A	High	6.52
rs1135071	HBB_R31S		R31S	High	8.77
rs1141387	HBB_V35F		V35F	Medium	5.66
rs1141387	HBB_V35L		V35L	Medium	4.33
rs11549407	HBB_Q40E	Uniprot residue:A	Q40E
rs33910209	HBB_Q132E		Q132E	High	9.23
rs33910209	HBB_Q132K		Q132K	High	7.33
rs33910569	HBB_Q128R		Q128R	High	6.56
rs33913712	HBB_E91K		E91K	Medium	3.45
rs33914944	HBB_R105S		R105S	Low	1.32
rs33915112	HBB_E27V		E27V	Medium	4.67
rs33915112	HBB_E27G		E27G	Medium	5.62
rs33915112	HBB_E27A	Uniprot residue:L	E27A
rs33916412	HBB_L29R		L29R	High	8.43
rs33916412	HBB_L29P		L29P	High	5.17
rs33916412	HBB_L29Q		L29Q	Medium	2.99
rs33917628	HBB_S90T		S90T	Low	1.33
rs33917628	HBB_S90N		S90N	Low	1.78
rs33917785	HBB_L92R		L92R	Medium	4.56
rs33917785	HBB_L92P	Uniprot residue:R	L92P
rs33918338	HBB_H144L		H144L	Low	0.99
rs33918338	HBB_H144R		H144R	Medium	2.33
rs33918338	HBB_H144P		H144P	Medium	4.98
rs33918343	HBB_V68G	Uniprot residue:W	V68G
rs33918343	HBB_V68A		V68A	High	6.77
rs33918343	HBB_V68E		V68E	Medium	4.32
rs33921821	HBB_A143V		A143V	High	12.33
rs33921821	HBB_A143D		A143D	High	11.34
rs33922842	HBB_E44Q		E44Q	Medium	9.87
rs33922842	HBB_E44K	Uniprot residue:V	E44K
rs33922873	HBB_H64Y		H64Y	Low	1.66
rs33922873	HBB_H64N		H64N	High	7.89
rs33924146	HBB_F43V	Uniprot residue:A	F43V
rs33924146	HBB_F43L		F43L	Medium	3.57
rs33924775	HBB_H93Y		H93Y	Medium	2.99
rs33924775	HBB_H93D		H93D	High	6.55
rs33924775	HBB_H93N	Uniprot residue:V	H93N
rs33925391	HBB_V127G		V127G	High	7.32
rs33925391	HBB_V127A		V127A	Medium	3.66
rs33925391	HBB_V127E		V127E	High	12.99
rs33926764	HBB_K9E		K9E	Low	0.78
rs33926764	HBB_K9Q	Uniprot residue:F	K9Q
rs33929459	HBB_V24F		V24F	Medium	4.44
rs33929459	HBB_V24I		V24I	Medium	5.01
rs33930165	HBB_E7Q		E7Q	High	9.23
rs33930165	HBB_E7K	Uniprot residue:H	E7K
rs33930702	HBB_M1I		M1I	High	7.23
rs33930702	HBB_M1I		M1I	Low	1.34

10. Active Site

Rank	C-Score	Cluster Size	PDB Hit	Lig Name	Consensus Building Residues
1	1.00	2019	1fsxA	HEM	43,45,64,67,68,71, 72,89,92,93,97,99, 103,104,107,138,142
2	0.06	71	4kmvA	T6C	28,29,32,64,65,68,69,107
3	0.04	34	3sdnA	ACT	43,64,68,107
4	0.03	24	3ic0A	B77	2,3,132,133,136,137,140
5	0.03	41	2oifB	CYN	29,64,68
6	0.02	41	3g4wA	8CL	19,24,28,69,72,111,114,115,131
7	0.02	17	2qmbD	OXY	64,68,93
8	0.02	15	1sdkD	TMM	2,3,83
9	0.01	20	2d2mD	k-mer	88,92
10	0.01	6	4ni0B	2P3	102,105,106,109,110,113

11. Primary Sequence Analysis

Number of amino acids

147

Molecular weight

15998.41

Theoretical pI

6.74

Amino acid composition

Ala (A) 15 10.2% Arg (R) 3 2.0%

Asn (N) 6 4.1%

Asp (D) 7 4.8%

Cys (C) 2 1.4%

Gln (Q) 3 2.0%

Glu (E) 8 5.4%

Gly (G) 13 8.8%

His (H) 9 6.1%

Ile (I) 0 0.0%

Leu (L) 18 12.2% Lys (K) 11 7.5%

Met (M) 2 1.4%

Phe (F) 8 5.4%

Pro (P) 7 4.8%

Ser (S) 5 3.4%

Thr (T) 7 4.8%

Trp (W) 2 1.4%

Tyr (Y) 3 2.0%

Val (V) 18 12.2% Pyl (O) 0 0.0%

Sec (U) 0 0.0%

Total number of negatively charged residues (Asp + Glu)

Total number of positively charged residues (Arg + Lys)

Formula

C₇₂₉H₁₁₂₈N₁₉₆O₂₀₂S₄

Total number of atoms(12269)

2259

Instability index(II)

6.16

Stability

stable

Aliphatic index(AI)

93.47

GRAVY

0.014

12. Secondary structure analysis

Alpha helix (Hh) : 102 is 69.39%

3₁₀helix (Gg) : 0 is 0.00%

Pi helix (Ii) : 0 is 0.00%

Beta bridge (Bb) : 0 is 0.00%

Extended strand (Ee) : 2 is 1.36%

Beta turn (Tt) : 0 is 0.00%

Bend region (Ss) : 0 is 0.00%

Random coil (Cc) : 43 is 29.25%

Ambiguous states (?) : 0 is 0.00%

Other states : 0 is 0.00%

Secondary Structure Prediction

Amino Acid Type

13. Enzymes

Name of enzyme	No. of cleavages
Arg-C proteinase	3
Asp-N endopeptidase	7
Asp-N endopeptidase + N-terminal Glu	15
BNPS-Skatole	2
CNBr	2
Chymotrypsin-high specificity (C-term to [FYW], not before P)	12
Chymotrypsin-low specificity (C-term to [FYWML], not before P)	41
Clostripain	3
Formic acid	7
Glutamyl endopeptidase	8
Iodosobenzoic acid	2
LysC	11
LysN	11
NTCB (2-nitro-5-thiocyanobenzoic acid)	2
Pepsin (pH1.3)	37
Pepsin (pH>2)	43
Proteinase K	79
Staphylococcal peptidase I	7
Thermolysin	54
Trypsin	14

14. Epitope

Rank	Start position	Sequence	Score	Prediction
1	8	EKSAVTALWGKVNVDE	0.85	Epitope
2	43	FESFGDLSTPDAVMGN	0.84	Epitope
3	37	PWTQRFFESFGDLSTP	0.83	Epitope
3	22	DEVGGEALGRLLVVYP	0.83	Epitope
4	119	FGKEFTPPVQAAYQKV	0.81	Epitope
5	90	SELHCDKLHVDPENFR	0.80	Epitope
6	125	PPVQAAYQKVVAGVAN	0.79	Epitope
7	52	PDAVMGNPKVKAHGKK	0.77	Epitope
8	58	NPKVKAHGKKVLGAFS	0.75	Epitope
9	110	VLVCVLAHHFGKEFTP	0.74	Epitope
10	74	DGLAHLDNLKGTFATL	0.73	Epitope
11	66	KKVLGAFSDGLAHLDN	0.72	Epitope
12	81	NLKGTFATLSELHCDK	0.64	Epitope
13	99	VDPENFRLLGNVLVCV	0.59	Epitope

15. Protein-Protein Interaction

No of Nodes	11
No of Edges	36
Avg node degree	6.55
avg. local clustering coefficient	0.845
expected number of e, dges	11
p-value	8.39e-10
Protein – Protein Interaction network

16. MRNA

RNA Base Pairing Probability Plot