. 2003 Mar;77(5):2866–2872. doi: 10.1128/JVI.77.5.2866-2872.2003

TABLE 1.

LCMV GP sequence at the cleavage site and results of mutational analysis

Expt	LCMV GP variant^a	Sequence at the cleavage site	Cleavage^b	Cell surface expression^c
Alanine scanning	Wild type	`KFLTRRLA₂₆₅ GTF₂₆₈`	+++	+++
Alanine scanning	K258A	`A..........`	+++	+++
	F259A	`.A.........`	−/+	+++
	L260A	`..A........`	+	+++
	T261A	`...A.......`	+	+++
	R262A	`....A......`	−	+++
	R263A	`.....A.....`	−	+++
	L264A	`......A....`	−	+++
	G266A	`........A..`	+++	+++
	T267A	`.........A.`	+++	+++
	F268A	`..........A`	+++	+++
Mutation at amino acids 262-264	Wild type	`KFLTRRLA₂₆₅ GTF₂₆₈`	+++	+++
	R262K	`....K......`	−	+++
	R263K	`.....K.....`	+++	+++
	R263H	`.....H.....`	++	+++
	R263S	`.....S.....`	+	+++
	R263N	`.....N.....`	+	+++
	L264I	`......I....`	+	+++
	L264V	`......V....`	−	+++
	L264F	`......F....`	−	+++
Mutation at amino acid 265	Wild type	`KFLTRRLA₂₆₅ GTF₂₆₈`	+++	+++
	A265L	`.......L...`	+++	+++
	A265I	`.......I...`	−	+++
	A265V	`.......V...`	−	+++
	A265T	`.......T...`	+++	+++
	A265F	`.......F...`	+++	+++
	A265E	`.......E...`	+	+++
	265S^d	`.......S...`	+++	+++

293T cells were transfected with expression plasmids encoding wild-type or mutated LCMV GPs with the indicated amino acid substitutions.

Cells were lysed 2 days after transfection, and the cleavage of each mutant relative to that of wild-type LCMV GP was determined by immunoblot analysis. − undetectable cleavage; +++, cleavage equivalent to that of wild-type LCMV GP; ++, good clevage but significant less cleavage product than for the wild-type LCMV GP; +, very inefficient but detectable cleavage; −/+, trace amounts of the cleavage product detectable in only two of three experiments.

Cells were stained 2 days after transfection with anti-LCMV GP-directed antibody KL25 and analyzed by flow cytometry. +++, cell surface expression at least equivalent to that of wild-type LCMV GP.

LCMV GP variant WE_P110L, which contains a serine at amino acid 265 (6).