Novel Antibacterial Activity of β2-Microglobulin in Human Amniotic Fluid 英文参考文献.docVIP

NovelAntibacterialActivityofb2-Microglobulinin HumanAmnioticFluid Jin-YoungKim1,Seong-CheolPark1,Jong-KookLee1,SangJoonChoi2,Kyung-SooHahm3, YoonkyungPark1,4 * 1ResearchCenterforProteinaceousMaterials,ChosunUniversity,Kwangju,Korea,2DepartmentofObstetricsandGynecology,SchoolofMedicine,ChosunUniversity, Kwangju,Korea,3BioleadersCorporation,Yusong-Ku,Daejeon,Korea,4DepartmentofBiotechnology,ChosunUniversity,Kwangju,Korea Abstract An antibacterial protein (about 12 kDa) was isolated from human amniotic fluid through dialysis, ultrafiltration and C18 reversed-phase HPLC steps. Automated Edman degradation showed that the N-terminal sequence of the antibacterial protein was NH2-Ile-Gln-Arg-Thr-Pro-Lys-Ile-Gln-Val-Tyr-Ser-Arg-His-Pro-Ala-Glu-Asn-Gly-. The N-terminal sequence of the antibacterialproteinwasfoundtobeidenticaltothatofb2-microglobulin,acomponentofMHCclassImolecules,whichare presentonallnucleatedcells.Matrix-assistedlaserdesorptionionizationmassspectrometry(MALDI-MS)revealedthatthe molecularmassoftheantibacterialproteinwas11,631Da.Thisantibacterialprotein,b2M,possessedpotentantibacterial activityagainstpathogenicbacteria.Specially,antibacterialactivitywasobservedinpotassiumbuffer,andpotassiumion was found to be critical for the antibacterial activity. Interestingly, the antibacterial action of b2M was associated with dissipationofthetransmembranepotential,buttheproteindidnotcausedamagetothemembranethatwouldresultin SYTOX green uptake. In addition, stimulation of WISH amniotic epithelial cells with the bacterial endotoxin lipopolysaccharide(LPS)induceddose-dependentupregulationofb2MmRNAexpression.Theseresultssuggestthatb2M contributestoaself-defenseresponsewhenamnioticcellsareexposedtopathogens. Citation:KimJ-Y,ParkS-C,LeeJ-K,ChoiSJ,HahmK-S,etal.(2012)NovelAntibacterialActivityofb2-MicroglobulininHumanAmnioticFluid.PLoSONE7(11): e47642.doi:10.1371/journal.pone.0047642 Editor:OlleTerenius,SwedishUniversityofAgriculturalSciences,Sweden ReceivedFebruary27,2012;