Novel Antibiotics Could Arise from Adhesion Protein Structure Analysis

Researchers at Harvard Medical School, the University of California-San Francisco, and the University of Georgia have described how the protein that allows strep and staph bacteria to stick to human cells is prepared and packaged.

Their study (“Unraveling the sequence of cytosolic reactions in the export of GspB adhesion from Streptococcus gordonii”), which could facilitate the development of new antibiotics, appears in the Journal of Biological Chemistry.

“Many pathogenic bacteria, including Streptococcus gordonii, possess a pathway for the cellular export of a single serine-rich-repeat protein that mediates the adhesion of bacteria to host cells and the extracellular matrix. This adhesin protein is O-glycosylated by several cytosolic glycosyltransferases and requires three accessory Sec proteins (Asp1–3) for export, but how the adhesin protein is processed for export is not well understood. Here, we report that the S. gordonii adhesin GspB is sequentially O-glycosylated by three enzymes (GtfA/B, Nss, and Gly) that attach N-acetylglucosamine and glucose to Ser/Thr residues,” wrote the investigators.

“We also found that modified GspB is transferred from the last glycosyltransferase to the Asp1/2/3 complex. Crystal structures revealed that both Asp1 and Asp3 are related to carbohydrate-binding proteins, suggesting that they interact with carbohydrates and bind glycosylated adhesin, a notion that was supported by further analyses. We further observed that Asp1 also has an affinity for phospholipids, which is attenuated by Asp2. In summary, our findings support a model in which the GspB adhesin is sequentially glycosylated by GtfA/B, Nss, and Gly and then transferred to the Asp1/2/3 complex in which Asp1 mediates the interaction of the Asp1/2/3 complex with the lipid bilayer for targeting of matured GspB to the export machinery.”

All bacteria have a standard secretion system that allows them to export different types of proteins outside of their cells. An important class of extracellular molecules produced by pathogenic bacteria is adhesins, proteins that enable bacteria to adhere to host cells. For unknown reasons, the SRR (serine-rich-repeat) adhesins of Staphylococcus and Streptococcus bacteria – athogens that can be involved in serious infections such as bacterial meningitis, bacterial pneumonia, and pericarditis – are transported via a secretion pathway that is similar to the standard system, but dedicated solely to adhesin.

Tom Rapoport, Ph.D., a professor at Harvard Medical School who oversaw the new study, wanted to understand what exactly these dedicated molecular supply chains were doing.

“I was intrigued by the fact that there is a second secretion system in some bacteria that is separate from the canonical secretion system and is just dedicated to the secretion of one protein,” Dr. Rapoport said. “There is a whole machinery and it’s only doing one thing.”

Yu Chen, Ph.D., at the time a postdoctoral research associate in Dr. Rapoport’s lab, led the investigation. She found that, in order to be transported, the adhesin protein needed to be modified with specific sugars by three enzymes acting in a specific sequence. These sugar modifications stabilize the protein and enhance its stickiness to target cells. Furthermore, the experiments showed that two proteins in the adhesin-specific pathway, whose function had previously been mysterious, seemed to be able to bind to these sugars, presumably enabling them to carry the adhesin to the cell membrane where adhesin’s dedicated exit channel is located.

The complexity of the adhesin transport system necessitated collaboration with research teams at UCSF, Harvard Medical School, and the University of Georgia.

“It’s a complicated system because it involves protein modification, chaperone activity, and membrane targeting, so we encountered a lot of challenges,” Dr. Chen said. “This [study] is a great example of how collaboration across labs in the scientific community advances human knowledge.”

The reason that these bacteria use this separate export pathway for adhesins remains elusive. But because this pathway is unique to strep and staph bacteria, the new understanding of its components could help researchers develop highly targeted antibiotics to treat infections caused by these bacteria in the future.

“You could imagine that you could develop novel antibiotics that could target this pathway,” according to Dr. Rapoport.