Architecture of the native major royal jelly protein 1 oligomer

Abstract

Honeybee caste development is nutritionally regulated by royal jelly (RJ). Major royal jelly protein 1 (MRJP1), the most abundant glycoprotein among soluble royal jelly proteins, plays pivotal roles in honeybee nutrition and larvae development, and exhibits broad pharmacological activities in humans. However, its structure has long remained unknown. Herein, we identify and report a 16-molecule architecture of native MRJP1 oligomer containing four MRJP1, four apisimin, and eight unanticipated 24-methylenecholesterol molecules at 2.65 Å resolution. MRJP1 has a unique six-bladed β-propeller fold with three disulfide bonds, and it interacts with apisimin mainly by hydrophobic interaction. Every four 24-methylenecholesterol molecules are packaged by two MRJP1 and two apisimin molecules. This assembly dimerizes to form an H-shaped MRJP14-apisimin4-24-methylenecholesterol8 complex via apisimin in a conserved and pH-dependent fashion. Our findings offer a structural basis for understanding the pharmacological effects of MRJPs and 24-methylenecholesterol, and provide insights into their unique physiological roles in bees.

The rise and fall of major royal jelly proteins during a honeybee (Apis mellifera) workers' life

“…As MRJP1 and apisimin form in RJ a complex with a stoichiometry of 4:4 (Mandacaru et al, ; Tian et al, ) and as relative transcript abundance is similar for both genes (Figure ), one would expect to quantify approximately the same molar amount of apisimin as MRJP1 at least in the HG at day 8. However, within the present study apisimin was not detected at all.…”

“…As MRJP1 and apisimin form in RJ a complex with a stoichiometry of 4:4 (Mandacaru et al, 2017;Tian et al, 2018) and as relative transcript abundance is similar for both genes (Figure S3) (Table S6). Within the brain, only MRJP1 was found within the 20 most abundant proteins at day 8 (37.8 ± 9.8 fmol; 5th place) (Table S6).…”

“…Because of the known complex formation of MRJP1 and apisimin in RJ with a stoichiometry of 4:4 (Mandacaru et al, 2017;Tian et al, 2018), we explicitly compared transcript abundance of these two genes which did not differ within the same day between days 0 and 20, but apisimin had significantly more transcripts than mrjp1 on day 24 in both brains and HGs (HG: 44-fold (p < 0.001); brain: 32-fold (p = 0.034)) (one-way ANOVA, p < 0.001, df = 27, F = 43.046; see Figure S3).…”

“…This fibrillary network confers the needed viscosity to royal jelly to prevent queen larvae falling out of their vertically oriented queen cells (Buttstedt et al, ; Kurth, Kretschmar, & Buttstedt, ). In addition, the complex of oligoMRJP1/apisimin binds 24‐methylenecholesterol and provides the developing larvae with essential sterols (Tian et al, ). MRJP3 binds and stabilizes RNA in royal jelly and is thought to share RNA among individuals (Maori et al, ).…”

Kikuji Yamaguchi Principles of Natural Beekeeping: A Novel Bio-Method of Natural Beekeeping for High Quality Royal Jelly Production

“…The author found that MRJP-1 multimer was stable at 2°C or À18°C. According to the most recent findings, the core structure of the MRJP1 multimer consists of four molecules of MRJP1, four molecules of the peptide apisimin [34] and, surprisingly, eight molecules of 24-methylenecholesterol [35,36].…”