Phenotypic and genetic analyses of four cases of coagulation factor XII deficiency

Figures & data

Table 1. Primer sequences, the length of amplification products and annealing temperature.

Exon	Primer sequences	Amplification products	Annealing temperature
E1 + 2	Forward Primer: 5′-AGGGCAGCTTGACCAATC-3′ Reverse Primer: 5′-GAGGAGCCAGGCCACTTA-3′	594 bp	60°C
E3 + 4	Forward Primer: 5′-GTGCCTGAGCAGTTGAGT-3′	437 bp	58°C
	Reverse Primer: 5′-AGTAATGAGGCGGGAGGA-3′
E5 + 6	Forward Primer: 5′-CTGCTCTTCCTTCCACCAT-3′	434 bp	60°C
	Reverse Primer: 5′-CCGTTCCCAACCATCTGC-3′
E7 + 8	Forward Primer: 5′-CACTGCGTTCCCTCCCCAAG-3′	600 bp	58°C
	Reverse Primer: 5′-CGTTAGAGCGCCGGGAG-3′
E9 + 10	Forward Primer: 5′-CCGTCGTGTGGCTACAGGA-3′	594 bp	58°C
	Reverse Primer: 5′-CTCTGGGCGGGCGGGTA-3′
E11 + 12	Forward Primer: 5′-GGGCTTCCCCAGAACGCTT-3′	493 bp	58°C
	Reverse Primer: 5′-TCTTCCGCCTAACCCAGTG-3′
E13 + 14	Forward Primer: 5′-GCGGCAAGCGTTGTCAGA-3′	639 bp	58°C
	Reverse Primer: 5′-GCGGGACCCATGAAGAAA-3′

Table 2. Hemostatic laboratory results.

Subject	PT(s)	APTT(s)	FIB(g/L)	FVIII:C(%)	FIX:C(%)	FXI:C(%)	FXII:C(%)	FXII:Ag(%)
Case 1	12.7	126.3	2.47	149	86	68	2	10
Case 2	13.1	122	3.16	124	129	83	1	11
Case 3	14	119	2.21	107	75	82	2	7
Case 4	13.1	126.5	3.2	187	101	70	2	6
Reference intervals	12.00–15.00	30.00–45.00	2.0–4.0	78–128	68–128	82–118	72–113	72–113

Figure 1. Information of mutant FXII DNA. (a1) NM_000505.4:c.398-1G > A in case 1. (a2) NC_000005.10:g.7217_7221delinsGTCTA in case 1. (b1) NM_000505.4:c.398-1G > A in case 2. (b2) NP_000496.2:p.(Ser479Ter) in case 2. (c1) NP_000496.2:p.(Pro182Leu) in case 3. (c2) NM_000505.4:c.1681-1G > A in case 3. (d1) NP_000496.2:p.(Cys559Arg) in case 4. (n) showed normal.

Table 3. Screening of the mutation site.

subject	Mutation	position	Style
Case 1	NM_000505.4:c.398-1G > A	Intron 5	Heterozygous mutation
	NC_000005.10:g.7217_7221delinsGTCTA	Exon 14
Case 2	NM_000505.4:c.398-1G > A	Intron 5	Heterozygous mutation
	NP_000496.2:p.(Ser479Ter)	Exon 12
Case 3	NP_000496.2:p.(Pro182Leu)	Exon 7	Heterozygous mutation
	NM_000505.4:c.1681-1G > A	Intron 13
Case 4	NP_000496.2:p.(Cys559Arg)	Exon 13	Homozygous mutation

Figure 2. Conserved analysis of missense mutation site. (a) 182 P- FXII’s conservatism as the black arrow showed. (b) 559 C- FXII’s conservatism as the black arrow showed.

Figure 3. Predictive analytics of FXII protein structure. (n1) Wild-type FXII protein’s cartoon model n2 In wild-type FXII protein’s cartoon model, yellow represents light chain and green represents heavy chain. (a1) Mutant FXII (NP_000496.2:p.(Pro182Leu)) protein’s cartoon model. (b1) Mutant FXII (NP_000496.2:p.(Cys559Arg)) protein’s cartoon model. (n3) Local structure model of 181 N- FXII and 182 P- FXII in wild-type FXII protein. (a2) 162 N- FXII and 182 L- FXII built hydrogen bond in mutant FXII (NP_000496.2:p.(Pro182Leu)) protein. (n4) Disulfide bond between 559-C FXII and 590-C FXII in wild-type FXII protein. (b2) Disulfide bond between 559 and 590 disappeared in mutant FXII (NP_000496.2:p.(Cys559Arg)) protein.