A. Rodrigue et al., REQUIREMENT FOR NICKEL OF THE TRANSMEMBRANE TRANSLOCATION OF NIFE-HYDROGENASE-2 IN ESCHERICHIA-COLI, FEBS letters, 392(2), 1996, pp. 81-86
The cellular location of membrane-bound NiFe-hydrogenase 2 (HYD2) from
Escherichia coli was studied by immunoblot analysis and by activity s
taining, Treatment of spheroplasts with trypsin was able to release ac
tive HYD2 into the soluble fraction, indicating that HYD2 is attached
to the periplasmic side of the cytoplasmic membrane and that HYD2 unde
rgoes a trans-membrane translocation during its biosynthesis. By using
a nik mutant deficient in the high affinity specific nickel transport
system, we show that the intracellular availability of nickel is esse
ntial for the processing of the large subunit and for the transmembran
e translocation of HYD2. We also demonstrate that the processing of th
e precursor, which is related with nickel incorporation, can occur in
the membrane-depleted soluble fraction and that it is associated with
the increase in resistance to proteolysis of the processed form of the
large subunit. The mechanism of the transmembrane translocation of HY
D2 is discussed.