SecA: Difference between revisions

[http://www.nature.com/nature/journal/v455/n7215/full/nature07335.html SecA] Crystallized complexes containing Bacillus subtilis SecA without its non-essential carboxy-terminal domain, and either Thermotoga maritima SecYE or Aquifex aeolicus SecYEG. These crystals diffracted X-rays to a maximum resolution of 6.2 Å and 7.5 Å, respectively. A higher resolution data set (4.5 Å) was obtained for a complex in which both partners were from T. maritima and the SecYEG complex was seleno-methionine (Se-Met) derivatized. All complexes were crystallized in the detergent Cymal-6 in the presence of ADP and BeFx. The structure of the complex of B. subtilis SecA and T. maritima SecYE was determined by molecular replacement with a B. subtilis SecA structure21 and served as an initial model for the other complexes. The building of a 4.5 Å resolution model of the T. maritima SecA–SecY complex was facilitated by the Se-Met positions (Supplementary Fig. 1), and by the high quality of the phases, leading to an electron density map that allowed the identification of large amino acid side chains (Fig. 1a and Supplementary Fig. 2). Model building also took into account conserved interactions between amino acids in previously determined SecA and SecY structures5, 21, 22 (sequence alignments are shown in Supplementary Figs 3 and 4). The final structure was refined to Rwork and Rfree factors of 27.9% and 30.3% (Table 1), respectively, and was used for all interpretations. It comprises all residues of SecA and most residues of SecYEG. No model could be built for the periplasmic loop between TM1 and TM2a of SecY (residues 42–61), as well as for residues of some termini (SecY residues 1–7 and 424–431; SecE residues 1–9; SecG residues 1–8 and 74–76). Furthermore, there are uncertainties about the tip of the loop between TM6 and TM7 (residues 240–254). An ADP–BeF3- complex was modelled into the electron density observed in the nucleotide-binding pocket of SecA (Supplementary Fig. 5). http://www.nature.com/nature/journal/v455/n7215/full/nature07335.html

==Function==

[http://journal.shouxi.net/qikan/article.php?id=418668 SecA] SecA interacts not only with the SecY channel (8) but also with acidic phospholipids (9-11) and with both the signal sequence and the mature part of a substrate protein (12). It also binds the chaperone SecB, which ushers some precursor proteins to SecA (8, 13, 14). When associated with the SecY complex, SecA undergoes repeated cycles of ATP-dependent conformational changes, which are linked to the movement of successive segments of a polypeptide chain through the channel (15, 16). However the mechanism employed by SecA to translocate substrates polypeptide chains through the SecY channel remains largely unknown.

An important issue concerning the function of SecA is its oligomeric state during translocation. SecA is a dimer in solution (17, 18), and previous work argued that this is its functional state (19). An x-ray structure of Bacillus subtilis SecA also indicates the existence of a dimer (7). However, recent evidence raises the possibility that SecA might actually function as a monomer; in solution, SecA dimers are in rapid equilibrium with monomers (20, 21). Although the equilibrium favors dimers, it is shifted almost completely toward monomers in the presence of membranes containing acidic phospholipids or upon binding to the SecY complex (21). A synthetic signal peptide had a similar effect, although this result is controversial (22). A monomeric derivative of SecA containing six point mutations retained some in vitro translocation activity (21), but the low level of translocation precluded any firm conclusion. In addition, the previous results do not exclude models in which SecA cycles between monomeric and oligomeric states during the translocation of a polypeptide chain (22, 23). Most importantly, the functional oligomeric state of SecA in vivo remains to be established. http://journal.shouxi.net/qikan/article.php?id=418668