Table 1.

Schematic presentation of the single alanine substitutions’ effect on expression and activity of GNIP1Aa protein variants

General description	Position of replacedresidue in GNIP1Aa	No. of affectedpositions	Proteinexpression	WCRactivity
Positive control	GNIP1Aa		++	++
Negative control	Plu-MACPF		++	−
Low expression	389, 407, 415, 420, 439	5	±	±
I. Complete knockouts	393, 395, 456	3	++	−
II. Reduced activity	332, 340, 349, 351, 352, 375, 377, 381, 383, 391, 403, 413, 443, 458, 519, 523, 529	17	++	+
III. Activity similar to the wild type GNIP1Aa	For the list see SI Appendix, Fig. S3 and Table S2	95	++	++

++, Expression of a target protein or WCR toxicity is similar to the wild-type GNIP1Aa, 4.00 stunt/97% mortality; stunt in the range of 3.75–4.00, 83–100% mortality; −, no activity or similar to the negative control, Plu-MACPF, 0.44 stunt/11% mortality; ±, the level of protein expression is clearly below the wild-type GNIP1Aa (SI Appendix, Fig. S3), resulting in no or low toxicity of tested samples (scores below 1.8 stunt/40% mortality); +, reduced activity (stunt in the range of 1.00–3.67, 24–92% mortality) at the level of protein expression similar to the wild-type GNIP1Aa.