Mining the Royal Jelly Proteins: Combinatorial Hexapeptide Ligand Library Significantly Improves the MS-Based Proteomic Identification in Complex Biological Samples

Eliza Matuszewska; Joanna Matysiak; Grzegorz Rosiński; Elżbieta Kędzia; Weronika Ząbek; Jarosław Zawadziński; Jan Matysiak

doi:10.3390/molecules26092762

Mining the Royal Jelly Proteins: Combinatorial Hexapeptide Ligand Library Significantly Improves the MS-Based Proteomic Identification in Complex Biological Samples

Molecules. 2021 May 7;26(9):2762. doi: 10.3390/molecules26092762.

Authors

Eliza Matuszewska¹, Joanna Matysiak², Grzegorz Rosiński³, Elżbieta Kędzia⁴, Weronika Ząbek⁵, Jarosław Zawadziński⁶, Jan Matysiak¹

Affiliations

¹ Department of Inorganic and Analytical Chemistry, Poznan University of Medical Sciences, Grunwaldzka 6 Street, 60-780 Poznań, Poland.
² Faculty of Health Sciences, Calisia University, Kaszubska 13 Street, 62-800 Kalisz, Poland.
³ Department of Animal Physiology and Development, Faculty of Biology, Adam Mickiewicz University in Poznan, Uniwersytetu Poznańskiego 6 Street, 61-614 Poznań, Poland.
⁴ Department of Innovative Biomaterials and Nanotechnologies, Institute of Natural Fibres and Medicinal Plants-National Research Institute, Wojska Polskiego 71B Street, 60-630 Poznań, Poland.
⁵ Department of Pediatric Gastroenterology and Metabolic Diseases, Poznan University of Medical Sciences, Szpitalna 27/33 Street, 60-572 Poznań, Poland.
⁶ Department of Physiotherapy, Poznan University of Medical Sciences, 28 Czerwca 1956 r. 135/147 Street, 61-545 Poznań, Poland.

Abstract

Royal jelly (RJ) is a complex, creamy secretion produced by the glands of worker bees. Due to its health-promoting properties, it is used by humans as a dietary supplement. However, RJ compounds are not fully characterized yet. Hence, in this research, we aimed to broaden the knowledge of the proteomic composition of fresh RJ. Water extracts of the samples were pre-treated using combinatorial hexapeptide ligand libraries (ProteoMiner^TM kit), trypsin-digested, and analyzed by a nanoLC-MALDI-TOF/TOF MS system. To check the ProteoMiner^TM performance in the MS-based protein identification, we also examined RJ extracts that were not prepared with the ProteoMiner^TM kit. We identified a total of 86 proteins taxonomically classified to Apis spp. (bees). Among them, 74 proteins were detected in RJ extracts pre-treated with ProteoMiner^TM kit, and only 50 proteins were found in extracts non-enriched with this technique. Ten of the identified features were hypothetical proteins whose existence has been predicted, but any experimental evidence proves their in vivo expression. Additionally, we detected four uncharacterized proteins of unknown functions. The results of this research indicate that the ProteoMiner^TM strategy improves proteomic identification in complex biological samples. Broadening the knowledge of RJ composition may contribute to the development of standards and regulations, enhancing the quality of RJ, and consequently, the safety of its supplementation.

Keywords: MALDI-TOF-MS; ProteoMinerTM; beehive product; proteins; proteomics; royal jelly.

MeSH terms

Fatty Acids / chemistry*
Insect Proteins / analysis*
Ligands
Mass Spectrometry*
Oligopeptides / analysis*
Proteomics*

Substances

Fatty Acids
Insect Proteins
Ligands
Oligopeptides
royal jelly

Grants and funding

2016/23/D/NZ7/03949/Narodowe Centrum Nauki