Table IV.

Arabidopsis proteins whose de novo synthesis was induced by α-amanitin (data from Figs. 6 and 7)

No.	Exp. MM	Exp. PI	Arabidopsis Protein Name	Cov.	Theo. MM	Theo. PI	AGI No.	Relative Abundanced
	kD			%	kD			Ama/H2O
69a	43.67	6.29	12S seed storage protein precursor	18	50.56	6.53	At1g03880	≥100
87a	18.43	6.36	β-Cruciferin 12S seed storage protein	33	20.72	6.36	At1g03880	34.5 ± 0.22
141a	29.35	7.42	α-Cruciferin 12S seed storage protein	21	29.23	6.49	At5g44120	31.4 ± 0.58
214b	27.14	6.10	α-Cruciferin 12S seed storage protein	23	29.86	6.60	At1g03880	18.6 ± 0.28
82a	33.89	6.24	α-Cruciferin 12S seed storage protein (fragment)	42	34.68	6.42	At4g28520	18.2 ± 0.62
246b	32.75	6.54	Globulin seed storage protein precursor (fragment)	14	55.06	6.64	At3g22640	10.7 ± 0.51
85a	27.20	6.50	α-Cruciferin 12S seed storage protein	32	29.86	6.60	At1g03880	8.7 ± 0.31
245b	29.95	6.53	α-Cruciferin 12S seed storage protein	16	29.86	6.60	At1g03880	4.1 ± 0.49
76a	23.94	5.58	α-Cruciferin 12S seed storage protein (fragment)	27	34.68	6.42	At4g28520	2.7 ± 0.51
84a	30.46	6.61	α-Cruciferin 12S seed storage protein	42	29.23	6.49	At5g44120	2.7 ± 0.02
25a	23.32	6.89	β-Cruciferin 12S seed storage protein (fragment)	43	21.20	6.19	At4g28520	2.6 ± 0.21
77a	25.42	5.78	α-Cruciferin 12S seed storage protein (fragment)	30	34.68	6.42	At4g28520	2.4 ± 0.33
80a	32.64	5.85	α-Cruciferin 12S seed storage protein (fragment)	33	34.68	6.42	At4g28520	2.0 ± 0.49
83a	34.35	6.42	α-Cruciferin 12S seed storage protein	33	34.68	6.42	At4g28520	2.0 ± 0.15
88a	22.82	8.68	α-Cruciferin 12S seed storage protein (fragment)	44	34.68	6.42	At4g28520	2.0 ± 0.13
255b	21.65	5.18	Dehydrin RAB18-related protein	Seqc	18.46	7.10	At5g66400	2.4 ± 0.19
254b	22.10	4.96	Dehydrin RAB18-related protein	Seqc	18.46	7.10	At5g66400	2.0 ± 0.06
190a	101.60	5.82	HSP101	13	101.29	5.81	At1g74310	≥100
137a	76.06	5.07	HSP70	8	71.10	5.14	At3g12580	2.0 ± 0.11
34b	13.32	6.02	Major latex protein	37	18.06	6.88	At1g14950	≥100
99a	13.32	6.28	Major latex protein	59	18.06	6.88	At1g14950	21.0 ± 0.47
37a	40.29	6.49	Cytosolic GAPDH	26	36.91	6.62	At3g04120	≥100
79a	27.24	5.20	F4I1.20 protein	26	23.67	4.86	At2g44390	≥100
96a	64.62	6.08	β-Glucosidase	15	60.01	6.20	At3g21370	≥100
116a	30.24	5.77	P1 clone:MRA19, Strong similarity to unknown protein T05029	33	28.78	5.92	At5g45690	74.1 ± 0.28
144a	15.87	6.82	Peptidyl-prolyl cis-trans isomerase	48	18.37	7.68	At4g38740	6.8 ± 0.48
174b	38.70	5.73	Cys synthase or O-acetylserine-thiol-lyase	9	33.80	5.90	At4g14880	6.1 ± 0.30
175a	37.85	5.71	Cys synthase or O-acetylserine-thiol-lyase	30	33.80	5.90	At4g14880	≥100
247b	34.90	6.56	Hydrolase	19	37.64	8.90	At4g39955	2.8 ± 0.26
264b	18.70	6.05	β-Cruciferin 12S seed storage protein	29	20.72	6.36	At1g03880	11.6 ± 0.44

H₂O, tt2-1 seeds incubated for 1 d in water; Ama, α-amanitin, and tt2-1 seeds incubated for 1 d in 500 μm α-amanitin as described in “Materials and Methods”; Cov., coverage; Exp., experimental; Seq, protein identified by Edman sequencing; Theo., theoretical.

^aListed proteins correspond to previously identified proteins (Gallardo et al., 2001, 2002a).

^bListed proteins correspond to proteins identified during this work.

^cFrom Edman sequencing, the sequence QNRPGGQATE was found for both protein nos. 254 and 255; BLAST analysis showed that this sequence is present in the Arabidopsis dehydrin related RAB18 protein At5g66400. Note that although these two proteins correspond to abundant proteins in 2D gels colored with the GelCode blue stain reagent (see “Materials and Methods”), they are poorly stained by silver nitrate.

^dData obtained from densitometric analysis of individual spots from 2D gels revealed by PhosphorImager analysis (see Fig. 6 for examples of in vivo protein synthesis of seeds labeled with [³⁵S]Met): normalized spot volume in the tt2-1 mutant seeds incubated for 1 d in 500 μm α-amanitin divided by the normalized spot volume in the tt2-1 mutant seeds incubated for 1 d in water ±sd, from three different gels and independent extractions; ≥100 means that the accumulation level of the corresponding protein in seeds incubated 1 d on water was close to background.