SlyD

Case study 1: Yaozhong Hu, 2017

The gene for a protein domain, derived from a tumor marker was expressed in E. coli strain BL21 (DE3). The recombinant protein was purified from cell lysates through immobilized metal affinity chromatography and size-exclusion chromatography. A contaminating bacterial protein was consistently co-purified, even using stringent washing solutions containing 50 or 100 mM imidazole. Followed by mass spectrometry, the researchers identified the bacterial contaminant as FKBP-type peptidyl-prolyl cis-trans isomerase (SlyD) from E. coli. Using their SlyD-specific Nb 5 they generated an immune-complex that could be removed either by immunocapturing or by size exclusion chromatography. Both methods allow us to prepare a recombinant protein sample where the SlyD contaminant was quantitatively eliminated.

Fig1. Occurrence of SlyD recognized by Nb5 in lysates of various E. coli strains and absence of cross-reactive proteins in mammalian HeLa and HEK2937T cell lysates..

Fig2. Removal of SlyD contaminant from recombinant protein preparations after immunoprecipitation with Nb5.

Case study 2: Kyung-Yeon Han, 2007

The proteome profile of Escherichia coli BL21(DE3) generated in response to heat shock stress was analyzed by two-dimensional electrophoresis (2-DE), wherein SlyD was identified. As a fusion partner, as well as solubility enhancer, SlyD facilitated folding and significantly increased the solubility of many aggregation-prone heterologous proteins in E. coli cytoplasm. SlyD was very effective in sequestering interactive surfaces of heterologous proteins associated with non-specific protein-protein interactions and the formation of inclusion bodies, most likely as a result of intrinsic folding efficiencies and/or chaperone-like activities. SlyD was also shown to be suitable for the production of a biologically active fusion mutant of Pseudomonas putida cutinase that is of considerable biotechnological and commercial interest.

Fig3. Result of SDS–PAGE analysis, showing the solubility of recombinant G-CSF released from SlyD by enterokinase treatment.

Fig4. Time-course variation in PNB (or PNP) concentration and initial maximum activity (μmol/ml/min) of PNB hydrolysis by cell-free supernatant from recombinant E. coli cell lysates containing the recombinant fusion mutant, SlyD::CUT..

SlyD involved in several pathways and played different roles in them. We selected most pathways SlyD participated on our site, such as , which may be useful for your reference. Also, other proteins which involved in the same pathway with SlyD were listed below. Creative BioMart supplied nearly all the proteins listed, you can search them on our site.

SlyD has several biochemical functions, for example, . Some of the functions are cooperated with other proteins, some of the functions could acted by SlyD itself. We selected most functions SlyD had, and list some proteins which have the same functions with SlyD. You can find most of the proteins on our site.

SlyD has direct interactions with proteins and molecules. Those interactions were detected by several methods such as yeast two hybrid, co-IP, pull-down and so on. We selected proteins and molecules interacted with SlyD here. Most of them are supplied by our site. Hope this information will be useful for your research of SlyD.