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Original Research
Characterization of salivary protein during ovulatory phase of
menstrual cycle through MALDITOF/MS
Alagendran S1,2,3, Saibaba G1, Muthukumar S1, Rajkumar R1, Guzman RG2, Archunan G1
Department of Animal
Science, Centre for Pheromone
Technology, Bharathidasan
University, Tiruchirappalli,
Tamil Nadu, India, 2Department
of Pharmacology and Physiology,
Neurosensorial Physiology
Laboratory, UNAM, AV.
Universidad, Mexico, 3Department
of Biochemistry, Periyar
University, Salem, TamilNadu,
India
1
Received
: 240512
Review completed : 121012
Accepted
: 221112
ABSTRACT
Context: Predicting ovulation is the basis on which the fertile period is determined. Nowadays
there are many methods available to detect the ovulatory period. Unfortunately, these methods
are not always effective for accurate detection of ovulation. Hence, an attempt was made
to detect ovulation through single dimension sodium dodecyl sulphate polyacrylamide gel
electrophoresis(SDSPAGE) analysis of protein with the help of saliva ferning.
Aims: The aim of this study was to determine the association of protein level with endogenous
reproductive hormone level across the menstrual cycle.
Settings and Design: Salivary protein and its confirmation were evaluated during menstrual
cycle followed by SDSPAGE and Mass spectrometry.
Statistical Method Used: The protein content present in saliva throughout menstrual cycle is
trail by SPSS statistical software version.
Materials and Methods: Salivary proteins were investigated serially during preovulatory,
ovulatory and postovulatory periods of normal menstrual cycle in eighteen healthy volunteers.
The samples were collected in three consecutive menstrual cycles. Salivary protein was estimated
and analyzed by single dimension SDSPAGE.
Results: The results revealed significant variations in protein concentrations during the menstrual
cycle. Protein levels were maximum during ovulation and minimum during postovulatory phase.
Further, single dimension SDSPAGE analysis showed seven different fractions of proteins is
from 1490 kilo Dalton(kDa) in the three phases of the menstrual cycle.
Conclusions: Among the proteins, 48 kDa protein was more predominantly exhibited during
ovulatory phase than pre and postovulatory phase. The present study indicates that the protein
level and the specific protein band (48 kDa) through MALDITOF MS analysis might serve as
an indicator for ovulation.
Key words: Human, matrixassisted laser desorption/ionizationmass spectorometry, menstrual
cycle, saliva, single dimension sodium dodecyl sulfate polyacrylamide gel electrophoresis
Website:
www.ijdr.in
PMID:
***
DOI:
10.4103/0970-9290.116669
157
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Alagendran, etal.
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Alagendran, etal.
Protein assay
Staining methods
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Alagendran, etal.
RESULTS
Protein content in saliva across normal menstrual cycle
MALDITOF/MS
DISCUSSION
This study reveals that the nature of saliva considerably varied
depending upon the reproductive period. Saliva samples
obtained during the ovulatory period revealed a clear ferning
pattern which was not observed in either the preovulatory
or the postovulatory period[Figure1]. Consequently, our
results suggest a strong relationship between salivary ferning
patterns and ovulatory phase of menstrual cycle which is
in agreement with earlier studies.[34] However, unlike what
was reported for human cervical mucus and bovine vaginal
mucus,[35,36] we did not find the appearance of ferning
immediately while observing the saliva under the microscope
160
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Alagendran, etal.
Figure 1: Salivary ferning pattern across menstrual cycle, (a) Preovulatory phase (6-12 days) fern like appearance was formed due to ESH
stimulation; (b) Ovulatory phase (13-14 days) Estrogen fluctuation stimulates with salts like sodium ions. Fern like crystals was formed; (c)
Postovulatory phase (15-26 days), Fern like formation was decline due to formation of Luteinising hormone
Figure 4: Mass spectrum of 48kDa protein (UDPNacetylglucosamine pyrophosphorylase). The sequence stretches that are covered by the
peptide ion signals (68% sequence coverage) in the mass spectrum are underlined in red color
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Alagendran, etal.
End
331
387
399
492
535
544
600
736
780
893
945
986
1029
1031
Observed
996.5
1104.6
1006.5
983.4
713.3
1074.9
787.4
880.4
1026.5
722.4
1003.5
1140.5
890.4
977.5
Expected
994.25
1103.57
1004.25
982.45
712.69
1073.4
786.94
879.48
1025.48
721.56
1002.3
1139.57
889.35
976.24
Sequence
MNINDLK.L*
M.NINDLKLTLSK.A*
K.AGQEHLLR.F
K.NVDARMEPVPR.E
R.MEPVPREVLGSATR.D
K.VAVLLLAGGQGTR.L*
K.GMYDVGLPSR.K
K.GMYDVGLPSRK.T
K.TLFQIQAER.I
K.CIIPWYIMTSGR.T
R.TMESTK.E
K.YFGLKK.E*
VADKEFHAPL*
IIDENGVHEL.VKNGI
*Matched peptides
ACKNOWLEDGMENTS
The study was partially supported by a grant from UGCSAP and
DSTFIST and DST-PURSE, New Delhi has kindly acknowledged.
Dr. SA was awarded by DGAPA Postdoctoral Research associate in
162
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How to cite this article: Alagendran S, Saibaba G, Muthukumar S, Rajkumar
R, Guzman RG, Archunan G. Characterization of salivary protein during
ovulatory phase of menstrual cycle through MALDI-TOF/MS. Indian J Dent
Res 2013;24:157-63.
Source of Support: Nil, Conflict of Interest: None declared.