In-Depth N-Glycosylation Reveals Species-Specific Modifications and Functions of the Royal Jelly Protein from Western (Apis mellifera) and Eastern Honeybees (Apis cerana).
Royal jelly (RJ), secreted by honeybee workers, plays diverse roles as nutrients and defense agents for honeybee biology and human health. Despite being reported to be glycoproteins, the glycosylation characterization and functionality of RJ proteins in different honeybee species are largely unknown. An in-depth N-glycoproteome analysis and functional assay of RJ produced by Apis mellifera lingustica (Aml) and Apis cerana cerana (Acc) were conducted. RJ produced by Aml yielded 80 nonredundant N-glycoproteins carrying 190 glycosites, of which 23 novel proteins harboring 35 glycosites were identified. For Acc, all 43 proteins glycosylated at 138 glycosites were reported for the first time. Proteins with distinct N-glycoproteomic characteristics in terms of glycoprotein species, number of N-glycosylated sites, glycosylation motif, abundance level of glycoproteins, and N-glycosites were observed in this two RJ samples. The fact that the low inhibitory efficiency of N-glycosylated major royal jelly protein 2 (MRJP2) against Paenibacillus larvae (P. larvae) and the absence of antibacterial related glycosylated apidaecin, hymenoptaecin, and peritrophic matrix in the Aml RJ compared to Acc reveal the mechanism for why the Aml larvae are susceptible to P. larvae, the causative agent of a fatal brood disease (American foulbrood, AFB). The observed antihypertension activity of N-glycosylated MRJP1 in two RJ samples and a stronger activity found in Acc than in Aml reveal that specific RJ protein and modification are potentially useful for the treatment of hypertensive disease for humans. Our data gain novel understanding that the western and eastern bees have evolved species-specific strategies of glycosylation to fine-tune protein activity for optimizing molecular function as nutrients and immune agents for the good of honeybee and influence on the health promoting activity for human as well. This serves as a valuable resource for the targeted probing of the biological functions of RJ proteins for honeybee and medical communities.
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Mao
M
Feng
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Yu
Y
Fang
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Bin
B
Han
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Xiang
X
Xu
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.|College of Bioengineering, Henan University of Technology , Zhengzhou 450001, China.
Pei
P
Fan
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Yue
Y
Hao
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Yuping
Y
Qi
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Han
H
Hu
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Xinmei
X
Huo
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Lifeng
L
Meng
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Bin
B
Wu
Institute of Apicultural Research/Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Chinese Academy of Agricultural Sciences , Beijing 100093, China.
Jianke
J
Li
D020816:Amino Acid Motifs; D000818:Animals; D000959:Antihypertensive Agents / Q000737:chemistry / Q000494:pharmacology; D001516:Bees / Q000737:chemistry*; D002478:Cells, Cultured; D005227:Fatty Acids / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry* / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000502:physiology; D040901:Proteomics / Q000379:methods; D016601:RNA-Binding Proteins; D013045:Species Specificity; D000099219:Royal Jelly
D000959:Antihypertensive Agents; D005227:Fatty Acids; D006020:Glycopeptides; D006023:Glycoproteins; D019476:Insect Proteins; D016601:RNA-Binding Proteins; D000099219:Royal Jelly; C559157:MRJP1 protein, Apis mellifera; C109593:MRJP3 protein, Apis mellifera
Apis cerana cerana; Apis mellifera ligustica; N-glycoproteome; Paenibacillus larvae; antihypertension; royal jelly