1. Supplementary figure 1:
spectroscopic analysis of the content of E. coli MC4100 cells carrying pTrc99A or a derivative recombinant plasmid
pTrc99A
pTrc99A-HasA(1-174)
pTrc99A-HasA(1-174)E148A, D167A 1

2. Supplementary figure 2:
Purification of (His)HasD by affinity chromatography MW
Western-blot analysis with anti-His tag antibodies
1: whole cell membranes solubilized in 0.7% lauryl maltoside
2: unbound fraction
3: washed fraction with 20 mM imidazole
4 and 5: eluted fraction with 250 mM imidazole
55 kDa
(His)HasD
Degradation products 2

3. Supplementary figure 3:
Secretion of HasA in cells expressing wild-type HasD or Walker B mutant HasDE474Q
HasA
(His)HasD
HasD WT E474Q
Coomassie-stained
Culture supernatants
WB His, HasA
Cell pellets
MC4100 ara pBAD24-(His)HasD-HasE, pTrc99A-HasA 3