Table 2 Immunocytochemical detection of AT/BT, FS, iGb3S, and GT activities.

From: Amino acid substitutions at sugar-recognizing codons confer ABO blood group system-related α1,3 Gal(NAc) transferases with differential enzymatic activity

Set	I				II				III				IV				V
Cell	HeLa(FUT2)				HeLa(FUT2)				COS1(B3GALNT1)				CHO (B3GALNT1 + GBGT1)				COS1
Antigen	A				B				FORS1				FORS1				α-Gal-epitope
Exp.	1	2	3	AT	1	2	3	BT	1	2	3	FS	1	2	3	iGb3S	1	2	3	GT
H_ABO-A
AGG	100.0	140.6	95.0	5+	0.0	0.0	0.0	—	16.7	15.0	15.7	3+	0.0	0.0	0.0	—	0.0	0.0	0.0	—
GGA	87.5	109.4	87.5	5+	44.0	35.7	36.7	4+	26.7	26.7	25.0	3+	0.0	0.0	0.0	—	0.0	0.0	0.0	—
HAA	0.3	0.6	0.5	+/—	48.0	39.3	40.0	4+	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—
LGG	100.0	100.0	100.0	5+	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—
MGA	2.5	3.1	3.0	+	100.0	100.0	100.0	5+	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—
M_GBGT1
AGG	0.5	0.7	0.4	+/−	3.2	2.1	1.7	+	90.0	83.3	83.3	5 +	0.0	0.0	0.0	—	0.0	0.0	0.0	—
GGA	3.8	3.4	3.3	+	0.8	1.1	0.3	+/−	100.0	100.0	100.0	5+	0.0	0.0	0.0	—	0.0	0.0	0.0	—
HAA	0.6	0.9	0.4	+/−	0.1	0.4	2.3	+/−	0.0	1.0	0.0	+/−	0.0	0.0	0.0	—	0.0	0.0	0.0	—
LGG	2.0	3.1	2.3	+	2.0	3.6	2.0	+	0.7	0.0	0.0	+/−	0.0	0.0	0.0	—	0.0	0.0	0.0	—
MGA	1.0	2.2	1.5	+/−	1.6	1.4	1.7	+/−	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—
R_A3GALT2
AGG	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	6.0	6.7	5.0	2+	0.0	0.0	0.0	—
GGA	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	5.0	5.6	6.0	2+	0.0	0.0	0.0	—
HAA	1.8	0.9	0.6	+/−	1.6	0.1	0.1	+/−	0.0	0.0	0.0	—	100.0	100.0	100.0	5+	4.2	2.5	5.0	+
LGG	0.3	0.0	0.1	—	0.6	0.5	0.3	+/−	0.0	0.0	0.0	—	22.0	22.2	20.0	3+	0.5	0.2	0.6	+/−
MGA	0.1	0.1	0.8	+/−	0.3	0.2	0.1	+/−	0.0	0.0	0.0	—	14.0	22.2	20.0	3+	0.0	0.3	0.0	—
M_GGTA1
AGG	0.0	0.1	0.3	—	0.2	0.0	0.3	+/−	0.0	0.0	0.0	—	0.8	0.0	0.0	+/−	0.2	0.5	0.6	+/−
GGA	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.2	0.2	0.2	+/−	16.7	16.7	16.0	3+
HAA	0.0	0.0	0.1	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.3	0.0	0.6	+/−	100.0	100.0	100.0	5+
LGG	0.0	0.0	0.0	—	0.1	0.0	0.0	—	0.0	0.0	0.0	—	0.2	0.2	0.4	+/−	1.7	0.8	2.0	+/−
MGA	0.0	0.2	0.0	—	0.0	0.0	0.0	—	0.0	0.0	0.0	—	0.6	0.7	0.6	+/−	11.7	10.0	10.0	2+

Five sets (I–V) of experiments were performed in triplicates (Exp. 1–3) using 4 cell types and 4 antigens analyzed. The original gene names and the tripeptide sequences at the codons corresponding to 266–268 of human AT/BT are listed in the leftmost column: H_ABO-A, M_GBGT1, R_A3GALT2, and M_GGTA1 for the A allele of human ABO gene, mouse GBGT1 gene, rat A3GALT2 gene, and mouse GGTA1 gene, respectively. The tripeptides are abbreviated: AGG (AlaGlyGly), GGA (GlyGlyAla), HAA (HisAlaAla), LGG (LeuGlyGly), and MGA (MetGlyAla). The original tripeptide sequences are shown in italic. The antigen positive cell percentages were re-calculated so that those of the original positive control constructs become 100%. The deduced AT, BT, FS, iGb3S, and GT activities are shown semi-quantitatively (-: no, +/−: scarce, + : very weak, 2 + : weak, 3 + : moderate, 4 + : strong, 5 + : very strong activity).

Back to article page

Table 2 Immunocytochemical detection of AT/BT, FS, iGb3S, and GT activities.

Search

Quick links