Table 1 The N-terminus of SecY contacts the C-terminus and the cytosolic loop C1 of YidC.

From: The interaction network of the YidC insertase with the SecYEG translocon, SRP and the SRP receptor FtsY

cross-link

cross-linked

SecY peptide

cross-linked

YidC peptide

calculated mass (Da)

mass deviation (ppm)

E-value

SecY(2)-YidC(545)

2AKQPGLDFQSAK13

539GLHSREKK546

2380.2862

2.5

0.0012

SecY(3)-YidC(413)

2AKQPGLDFQSAK13

402QRISQEMMALYKAEK416

3251.6681

0.6

0.0247

  1. E. coli BL21 cells carrying pTrc99a-SecYHisEG was used for SecYHisEG purification via metal affinity chromatography. The amine-specific cross-linker DSS was either added before SecYHisEG elution (on-column cross-linking) or after elution with elution buffer (solution cross-linking). Subsequently, samples were subjected to HPLC-MS. Cross-linked amino acids in the respective peptides are underlined and numbers in subscript indicate the position of the identified peptide within the protein. Identification of cross-linked peptides was performed essentially as described77 using the program pLink®79 for searching inter- or intra-molecular cross-links generated by amine-specific cross-linker DSS between peptides from SecY and YidC. The resulting set of peptide spectrum matches at a false discovery rate <0.05 was further filtered requiring a minimum E-value of 0.05 and identification of a pair of cross-linked residues in at least two independent experiments.
Back to article page