PMID- 10446356
OWN - NLM
STAT- MEDLINE
DCOM- 19991001
LR  - 20190610
IS  - 0006-3002 (Print)
IS  - 0006-3002 (Linking)
VI  - 1433
IP  - 1-2
DP  - 1999 Aug 17
TI  - Insect immune activation by recombinant Galleria mellonella apolipophorin III(1).
PG  - 16-26
AB  - Apolipophorin III (apoLp-III) is an exchangeable insect apolipoprotein. Its
      function, as currently understood, lies in the stabilization of low-density
      lipophorin particles (LDLp) crossing the hemocoel in phases of high energy
      consumption to deliver lipids from the fat body to the flight muscle cells.
      Recent studies with native Galleria mellonella-apoLp-III gave first indications
      of an unexpected role of that protein in insect immune activation. Here we report
      the immune activation by the recombinant protein, documenting a newly discovered 
      correlation between lipid physiology and immune defense in insects. The complete 
      cDNA sequence of G. mellonella-apoLp-III was identified by mixed
      oligonucleotide-primed amplification of cDNA (MOPAC), 3'-RACE-PCR, and cRACE-PCR.
      The sequence coding for the native protein was ligated into a pET-vector; this
      construct was transfected into Escherichia coli and overexpressed in the
      bacteria. Photometric turbidity assays with human low density lipoprotein (LDL)
      and transmission electron microscopy studies on apoLp-III-stabilized lipid discs 
      revealed the full functionality of the isolated recombinant apoLp-III with regard
      to its lipid-association ability. For proving its immune-stimulating capacity,
      apoLp-III was injected into the hemocoel of last instar G. mellonella larvae and 
      the antibacterial activity in cell-free hemolymph was determined 24 h later. As a
      result, the hemolymph samples of injected insects contained strongly increased
      antibacterial activities against E. coli as well as clearly enhanced
      lysozyme-like activities. From Northern blot analysis of total RNA from insects
      injected with apoLp-III or the bacterial immune provocator lipopolysaccharide, it
      could be concluded that the transcription rate of apoLp-III mRNA does not vary in
      comparison to untreated last instar larvae.
FAU - Niere, M
AU  - Niere M
AD  - Free University of Berlin, Institute of Zoology, Konigin-Luise-Str. 1-3, D-14195,
      Berlin, Germany.
FAU - Meisslitzer, C
AU  - Meisslitzer C
FAU - Dettloff, M
AU  - Dettloff M
FAU - Weise, C
AU  - Weise C
FAU - Ziegler, M
AU  - Ziegler M
FAU - Wiesner, A
AU  - Wiesner A
LA  - eng
SI  - GENBANK/AJ006975
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Biochim Biophys Acta
JT  - Biochimica et biophysica acta
JID - 0217513
RN  - 0 (Apolipoproteins)
RN  - 0 (DNA, Complementary)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Lipoproteins, LDL)
RN  - 0 (Recombinant Proteins)
RN  - 0 (apolipophorin III)
RN  - U86ZGC74V5 (Dimyristoylphosphatidylcholine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Apolipoproteins/chemistry/genetics/*pharmacology
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - DNA, Complementary/chemistry/metabolism
MH  - Dimyristoylphosphatidylcholine/chemistry
MH  - Escherichia coli/metabolism
MH  - Insecta/*drug effects/immunology/ultrastructure
MH  - Lipopolysaccharides/pharmacology
MH  - Lipoproteins, LDL/chemistry
MH  - Molecular Sequence Data
MH  - Recombinant Proteins/pharmacology
EDAT- 1999/08/14 00:00
MHDA- 1999/08/14 00:01
CRDT- 1999/08/14 00:00
PHST- 1999/08/14 00:00 [pubmed]
PHST- 1999/08/14 00:01 [medline]
PHST- 1999/08/14 00:00 [entrez]
AID - S016748389900148X [pii]
AID - 10.1016/s0167-4838(99)00148-x [doi]
PST - ppublish
SO  - Biochim Biophys Acta. 1999 Aug 17;1433(1-2):16-26. doi:
      10.1016/s0167-4838(99)00148-x.