Background
Methods
Cell culture
Royal jelly sample
Fractionation of royal jelly
Electrophoretic analysis of fractions
MTT cell viability assay
Scratch-wound assays
Identification and quantitative analysis of fractions possessing bioactivity
Functional analysis of the proteins identified in bioactive fractions
Statistical analysis
Results
Isolation and electrophoretic analysis of proteins from royal jelly
Proliferative effect of protein fractions on human epidermal keratinocytes
Cell migratory effects of protein fractions in an in vitro scratch wound model
Identification and quantification of bioactive protein fractions
Accession | Description | Abundances | Percentages |
---|---|---|---|
A0A1Q1N6G0 | Major royal jelly protein 2 OS = Apis mellifera carnica GN = MRJP2 PE = 4 SV = 1 | 12095077555 | 56.443% |
D3Y5T0 | Major royal jelly protein OS = Apis mellifera GN = MRJP3 PE = 2 SV = 1 | 6424005583 | 29.979% |
O77061 | Major royal jelly protein 2 OS = Apis mellifera GN = MRJP2 PE = 1 SV = 1 | 788699090.3 | 3.681% |
A0A087ZRA1 | Uncharacterized protein OS = Apis mellifera PE = 3 SV = 1 | 540461377.8 | 2.522% |
Q6IMJ9 | Major royal jelly protein 7 OS = Apis mellifera GN = MRJP7 PE = 2 SV = 1 | 345406314 | 1.612% |
O18330 | Major royal jelly protein 1 OS = Apis mellifera GN = MRJP1 PE = 1 SV = 1 | 202611735 | 0.946% |
A0A087ZW88 | Uncharacterized protein OS = Apis mellifera PE = 4 SV = 1 | 174868784 | 0.816% |
A0A087ZQI5 | ATP synthase subunit alpha OS = Apis mellifera GN = Atp5a1 PE = 3 SV = 1 | 130414480 | 0.609% |
A0A087ZRE3 | Elongation factor 1-alpha OS = Apis mellifera GN = EF1a-F2 PE = 3 SV = 1 | 117294891.1 | 0.547% |
A0A088A436 | Tubulin alpha chain OS = Apis mellifera GN = LOC550827 PE = 3 SV = 1 | 113409161.6 | 0.529% |
A0A088AN20 | Uncharacterized protein OS = Apis mellifera GN = RpL40 PE = 4 SV = 1 | 65715884.16 | 0.307% |
Q4ZJX1 | Major royal jelly protein 9 OS = Apis mellifera GN = MRJP9 PE = 2 SV = 1 | 58901241 | 0.275% |
A0A087ZQ27 | Uncharacterized protein OS = Apis mellifera PE = 3 SV = 1 | 44876322.19 | 0.209% |
A0A088AMB8 | ATP synthase subunit beta OS = Apis mellifera GN = Atp5b PE = 3 SV = 1 | 30241432 | 0.141% |
A0A088AEZ4 | Tubulin alpha chain OS = Apis mellifera GN = LOC552766 PE = 3 SV = 1 | 24948560 | 0.116% |
A0A088A5A6 | Uncharacterized protein OS = Apis mellifera GN = LOC409481 PE = 3 SV = 1 | 20733956.13 | 0.097% |
W8S9B2 | Actin (Fragment) OS=Nosema ceranae PE = 3 SV = 1 | 19727244.75 | 0.092% |
H9KL77 | Histone H4 OS = Apis mellifera GN = LOC724757 PE = 3 SV = 1 | 17945946.88 | 0.084% |
A0A087ZNX0 | Uncharacterized protein OS = Apis mellifera GN = Rab11 PE = 4 SV = 1 | 16626939 | 0.078% |
A0A087ZSC1 | Tubulin beta chain OS = Apis mellifera GN = LOC410559 PE = 3 SV = 1 | 14751532.47 | 0.069% |
A0A088AJJ6 | Uncharacterized protein OS = Apis mellifera GN = LOC411989 PE = 3 SV = 1 | 14168904 | 0.066% |
A0A088AGJ8 | Uncharacterized protein OS = Apis mellifera GN = LOC410620 PE = 3 SV = 1 | 11988349 | 0.056% |
A0A088A5X7 | Uncharacterized protein OS = Apis mellifera GN = LOC409167 PE = 4 SV = 1 | 11909793.5 | 0.056% |
A0A088A3F4 | Uncharacterized protein OS = Apis mellifera GN = LOC727045 PE = 4 SV = 1 | 11490301.06 | 0.054% |
A0A088A2A5 | Uncharacterized protein OS = Apis mellifera GN = LOC552272 PE = 3 SV = 1 | 11100238 | 0.052% |
A0A087ZUL8 | Uncharacterized protein OS = Apis mellifera GN = RpS15 PE = 3 SV = 1 | 10443703 | 0.049% |
A0A087ZR05 | Uncharacterized protein OS = Apis mellifera GN = Rpn11 PE = 4 SV = 1 | 9778002 | 0.046% |
A0A087ZV73 | Uncharacterized protein OS = Apis mellifera GN = LOC724873 PE = 4 SV = 1 | 8411322 | 0.039% |
A0A087ZMS7 | Uncharacterized protein OS = Apis mellifera GN = Rab39 PE = 4 SV = 1 | 7275442.125 | 0.034% |
A0A088A6D6 | Uncharacterized protein OS = Apis mellifera GN=Ndufs3 PE = 3 SV = 1 | 6409224.875 | 0.030% |
A0A088A7D1 | Uncharacterized protein OS = Apis mellifera GN = LOC410306 PE = 3 SV = 1 | 6343857.5 | 0.030% |
A0A088A8F0 | Putative H3K9 methyltransferase OS = Apis mellifera GN = 685996 PE = 4 SV = 1 | 6048775.25 | 0.028% |
A0A087ZYZ1 | Tubulin beta chain OS = Apis mellifera GN = LOC408782 PE = 3 SV = 1 | 6014038.938 | 0.028% |
A0A0B4J2N0 | Uncharacterized protein OS = Apis mellifera GN = LOC550794 PE = 3 SV = 1 | 4973486.5 | 0.023% |
A0A0B4J2L4 | Uncharacterized protein OS = Apis mellifera GN = LOC410026 PE = 3 SV = 1 | 4117948.25 | 0.019% |
A0A088AIY2 | Uncharacterized protein OS = Apis mellifera GN = mago PE = 4 SV = 1 | 4084139.625 | 0.019% |
A0A088AFT2 | 40S ribosomal protein S6 OS = Apis mellifera GN = LOC725647 PE = 3 SV = 1 | 3911799.438 | 0.018% |
A0A088ATI7 | Uncharacterized protein OS = Apis mellifera GN = LOC409126 PE = 4 SV = 1 | 3841522.813 | 0.018% |
A0A087ZMT8 | Uncharacterized protein OS = Apis mellifera PE = 4 SV = 1 | 3634688.25 | 0.017% |
A0A087ZZN8 | Proteasome subunit alpha type OS = Apis mellifera GN=Prosalpha5 PE = 3 SV = 1 | 3591316 | 0.017% |
A0A087ZV06 | 40S ribosomal protein S8 OS = Apis mellifera GN = Rps8 PE = 3 SV = 1 | 3381020.75 | 0.016% |
A0A088A6T4 | Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial OS = Apis mellifera GN=SdhA PE = 3 SV = 1 | 3201888.25 | 0.015% |
A0A088ADQ6 | Uncharacterized protein OS = Apis mellifera PE = 3 SV = 1 | 2594249.063 | 0.012% |
A0A088A2I2 | UDP-glucose 6-dehydrogenase OS = Apis mellifera GN = LOC413356 PE = 3 SV = 1 | 2218352.25 | 0.010% |
A0A088A9V8 | Uncharacterized protein OS = Apis mellifera PE = 3 SV = 1 | 2212138 | 0.010% |
A0A087EPB0 | Cell division protein FtsZ OS = Lactobacillus kunkeei GN = ftsZ PE = 3 SV = 1 | 2057526.938 | 0.010% |
A0A088ANZ0 | Uncharacterized protein OS = Apis mellifera GN = LOC551093 PE = 3 SV = 1 | 1902225 | 0.009% |
A0A088ARA9 | 60S ribosomal protein L13 OS = Apis mellifera GN = RpL13 PE = 3 SV = 1 | 1656290.5 | 0.008% |
A0A087EQF2 | 6-phosphogluconate dehydrogenase, decarboxylating OS = Lactobacillus kunkeei GN = JI66_01835 PE = 3 SV = 1 | 1644316.875 | 0.008% |
A0A088A2L4 | Uncharacterized protein OS = Apis mellifera PE = 3 SV = 1 | 1338471.375 | 0.006% |
V5 T859 | Glyceraldehyde-3-phosphate dehydrogenase OS=Bifidobacterium sp. Bin2N PE = 3 SV = 1 | 1259643.5 | 0.006% |
A0A088AEV2 | Uncharacterized protein OS = Apis mellifera GN = RpL26 PE = 4 SV = 1 | 1221172 | 0.006% |
A0A087ZW54 | Elongation factor Tu OS = Apis mellifera GN = LOC408328 PE = 3 SV = 1 | 1219225.375 | 0.006% |
A0A088AFM4 | Uncharacterized protein OS = Apis mellifera GN = TER94 PE = 3 SV = 1 | 1214927.25 | 0.006% |
D3JZ08 | MRJP5 OS = Apis mellifera PE = 2 SV = 1 | 802286.1875 | 0.004% |
A0A0B4J2P2 | Uncharacterized protein OS = Apis mellifera GN = LOC551386 PE = 3 SV = 1 | 788959.9375 | 0.004% |
A0A088ATP8 | Tubulin alpha chain OS = Apis mellifera PE = 3 SV = 1 | 654735.875 | 0.003% |
A0A087ZUP0 | Uncharacterized protein OS = Apis mellifera GN = 14–3-3epsilon PE = 3 SV = 1 | 638754.125 | 0.003% |
A0A088AJ01 | Mitogen-activated protein kinase OS = Apis mellifera GN = rl PE = 4 SV = 1 | 616170.75 | 0.003% |
A0A087ZMS5 | Uncharacterized protein OS = Apis mellifera PE = 4 SV = 1 | 579576 | 0.003% |
A0A087EQE2 | S-adenosylmethionine synthase OS = Lactobacillus kunkeei GN = metK PE = 3 SV = 1 | 554618.25 | 0.003% |
A0A088A9W4 | Uncharacterized protein OS = Apis mellifera GN=Flo1 PE = 3 SV = 1 | 333982.2188 | 0.002% |
A0A088ANC5 | APD-3-like protein; Apidermin 1-like protein; Apidermin 3-like protein OS = Apis mellifera GN = apd-3 PE = 4 SV = 1 | 332523.6875 | 0.002% |
Bioinformatic analysis of the proteins identified in fraction 2
Protein ID | Identity | E value | COG gene ID | COG num | Functional description | Functional class | Class description |
---|---|---|---|---|---|---|---|
A0A088A2A5 | 0.62 | 8.00E-51 | YP_875383 | COG0048 | Ribosomal protein S12 | J | Translation, ribosomal structure and biogenesis; |
A0A088A2I2 | 0.64 | 0 | YP_003548756 | COG1004 | UDP-glucose 6-dehydrogenase | M | Cell wall/membrane/envelope biogenesis; |
A0A088A2L4 | 0.43 | 0 | YP_634186 | COG0326 | Molecular chaperone, HSP90 family | O | Posttranslational modification, protein turnover, chaperones; |
A0A088A3F4 | 0.32 | 1.00E-05 | YP_002346601 | COG1530 | Ribonuclease G or E | J | Translation, ribosomal structure and biogenesis; |
A0A088A461 | 0.3 | 3.00E-17 | YP_001958455 | COG1196 | Chromosome segregation ATPase | D | Cell cycle control, cell division, chromosome partitioning; |
A0A088A5A6 | 0.41 | 6.00E-37 | YP_001737337 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
A0A088A5X7 | 0.34 | 1.00E-159 | YP_001736560 | COG0480 | Translation elongation factor EF-G, a GTPase | J | Translation, ribosomal structure and biogenesis; |
A0A088A6D6 | 0.52 | 8.00E-69 | YP_422148 | COG0852 | NADH:ubiquinone oxidoreductase 27 kD subunit (chain C) | C | Energy production and conversion; |
A0A088A6T4 | 0.64 | 0 | YP_742173 | COG1053 | Succinate dehydrogenase/fumarate reductase, flavoprotein subunit | C | Energy production and conversion; |
A0A088A7D1 | 0.38 | 1.00E-34 | YP_001737337 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
A0A088A8F0 | 0.49 | 1.00E-113 | YP_004484975 | COG5257 | Translation initiation factor 2, gamma subunit (eIF-2gamma; GTPase) | J | Translation, ribosomal structure and biogenesis; |
A0A088A9V8 | 0.67 | 1.00E-178 | YP_004616696 | COG1089 | GDP-D-mannose dehydratase | M | Cell wall/membrane/envelope biogenesis; |
A0A088ADQ6 | 0.6 | 2.00E-47 | YP_686965 | COG0100 | Ribosomal protein S11 | J | Translation, ribosomal structure and biogenesis; |
A0A088AEV2 | 0.46 | 2.00E-29 | NP_579542 | COG0198 | Ribosomal protein L24 | J | Translation, ribosomal structure and biogenesis; |
A0A088AFM4 | 0.48 | 0 | YP_004483987 | COG1222 | ATP-dependent 26S proteasome regulatory subunit | O | Posttranslational modification, protein turnover, chaperones; |
A0A087EPB0 | 0.72 | 0 | YP_007414867 | COG0206 | Cell division GTPase FtsZ | D | Cell cycle control, cell division, chromosome partitioning; |
A0A087EQE2 | 0.76 | 0 | NP_814529 | COG0192 | S-adenosylmethionine synthetase | H | Coenzyme transport and metabolism; |
A0A088AFT2 | 0.37 | 5.00E-19 | YP_001737450 | COG2125 | Ribosomal protein S6E (S10) | J | Translation, ribosomal structure and biogenesis; |
A0A088AGJ8 | 0.5 | 0 | NP_927210 | COG0443 | Molecular chaperone DnaK (HSP70) | O | Posttranslational modification, protein turnover, chaperones; |
A0A088AJ01 | 0.3 | 6.00E-24 | YP_008152560 | COG0515 | Serine/threonine protein kinase | T | Signal transduction mechanisms; |
A0A088AJJ6 | 0.42 | 1.00E-105 | YP_003640197 | COG0513 | Superfamily II DNA and RNA helicase | L | Replication, recombination and repair; |
A0A088AMB8 | 0.81 | 0 | YP_004357703 | COG0055 | FoF1-type ATP synthase, beta subunit | C | Energy production and conversion; |
A0A088AN20 | 0.84 | 6.00E-39 | YP_004089966 | COG5272 | Ubiquitin | O | Posttranslational modification, protein turnover, chaperones; |
A0A088ANZ0 | 0.54 | 0 | NP_276090 | COG1155 | Archaeal/vacuolar-type H + -ATPase catalytic subunit A/Vma1 | C | Energy production and conversion; |
A0A088ARA9 | 0.38 | 5.00E-07 | YP_006863204 | COG4352 | Ribosomal protein L13E | J | Translation, ribosomal structure and biogenesis; |
A0A088ATI7 | 0.34 | 8.00E-27 | YP_008431664 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
A0A0B4J2L4 | 0.5 | 1.00E-123 | NP_070800 | COG1222 | ATP-dependent 26S proteasome regulatory subunit | O | Posttranslational modification, protein turnover, chaperones; |
A0A0B4J2N0 | 0.52 | 1.00E-127 | NP_614161 | COG1222 | ATP-dependent 26S proteasome regulatory subunit | O | Posttranslational modification, protein turnover, chaperones; |
A0A0B4J2P2 | 0.54 | 1.00E-125 | NP_275871 | COG1222 | ATP-dependent 26S proteasome regulatory subunit | O | Posttranslational modification, protein turnover, chaperones; |
A0A1Q1N6G0
|
0.28
|
1.00E-17
|
YP_004643033
|
COG3386
|
Sugar lactone lactonase YvrE
|
G
|
Carbohydrate transport and metabolism;
|
D3JZ08 | 0.37 | 2.00E-16 | YP_001538809 | COG1158 | Transcription termination factor Rho | K | Transcription; |
D3Y5T0
|
0.3
|
4.00E-25
|
YP_004643033
|
COG3386
|
Sugar lactone lactonase YvrE
|
G
|
Carbohydrate transport and metabolism;
|
A0A087EQF2 | 0.77 | 0 | YP_007414230 | COG0362 | 6-phosphogluconate dehydrogenase | G | Carbohydrate transport and metabolism; |
A0A087ZMS7 | 0.34 | 9.00E-19 | YP_002463522 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
O18330 | 0.28 | 9.00E-20 | YP_004643033 | COG3386 | Sugar lactone lactonase YvrE | G | Carbohydrate transport and metabolism; |
O77061
|
0.28
|
1.00E-17
|
YP_004643033
|
COG3386
|
Sugar lactone lactonase YvrE
|
G
|
Carbohydrate transport and metabolism;
|
Q4ZJX1 | 0.3 | 3.00E-21 | YP_004643033 | COG3386 | Sugar lactone lactonase YvrE | G | Carbohydrate transport and metabolism; |
Q6IMJ9
|
0.3
|
2.00E-20
|
YP_004643033
|
COG3386
|
Sugar lactone lactonase YvrE
|
G
|
Carbohydrate transport and metabolism;
|
V5 T859 | 0.79 | 0 | YP_003986273 | COG0057 | Glyceraldehyde-3-phosphate dehydrogenase/erythrose-4-phosphate dehydrogenase | G | Carbohydrate transport and metabolism; |
W8S9B2 | 0.37 | 3.00E-70 | YP_003266114 | COG5277 | Actin-related protein | Z | Cytoskeleton; |
A0A087ZMT8 | 0.63 | 3.00E-13 | YP_008432142 | COG0638 | 20S proteasome, alpha and beta subunits | O | Posttranslational modification, protein turnover, chaperones; |
A0A087ZNX0 | 0.35 | 1.00E-29 | YP_008431664 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
A0A087ZQ27 | 0.51 | 0 | YP_001369341 | COG0443 | Molecular chaperone DnaK (HSP70) | O | Posttranslational modification, protein turnover, chaperones; |
A0A087ZQI5 | 0.73 | 0 | YP_423504 | COG0056 | FoF1-type ATP synthase, alpha subunit | C | Energy production and conversion; |
A0A087ZR05 | 0.35 | 1.00E-17 | YP_008797954 | COG1310 | Proteasome lid subunit RPN8/RPN11, contains Jab1/MPN domain metalloenzyme (JAMM) motif | O | Posttranslational modification, protein turnover, chaperones; |
A0A087ZRA1 | 0.37 | 1.00E-93 | YP_003266114 | COG5277 | Actin-related protein | Z | Cytoskeleton; |
A0A087ZRE3 | 0.55 | 0 | YP_003669641 | COG5256 | Translation elongation factor EF-1alpha (GTPase) | J | Translation, ribosomal structure and biogenesis; |
A0A087ZUL8 | 0.51 | 8.00E-43 | YP_003650076 | COG0185 | Ribosomal protein S19 | J | Translation, ribosomal structure and biogenesis; |
A0A087ZV06 | 0.41 | 8.00E-14 | YP_001030947 | COG2007 | Ribosomal protein S8E | J | Translation, ribosomal structure and biogenesis; |
A0A087ZV73 | 0.33 | 9.00E-30 | YP_008431664 | COG1100 | GTPase SAR1 family domain | R | General function prediction only; |
A0A087ZW54 | 0.57 | 1.00E-166 | YP_001995997 | COG0050 | Translation elongation factor EF-Tu, a GTPase | J | Translation, ribosomal structure and biogenesis; |
A0A087ZW88 | 0.35 | 3.00E-60 | YP_007146068 | COG3325 | Chitinase, GH18 family | G | Carbohydrate transport and metabolism; |
A0A087ZZN8 | 0.42 | 4.00E-62 | YP_003434754 | COG0638 | 20S proteasome, alpha and beta subunits | O | Posttranslational modification, protein turnover, chaperones; |