Serum concentration and interaction properties of MBL/ficolin associated protein-1

Mikkel-ole Skjoedt, Tina Hummelshoj, Yaseelan Palarasah, Estrid Hein, Lea Munthe-fog, Claus Koch, Karsten Skjodt, Peter Garred

19 Citations (Scopus)

Abstract

Recently, a novel protein named MBL/ficolin associated protein-1 (MAP-1) derived from the MASP1 gene through differential splicing was identified. In the present study, we established biochemical characteristics, determined the serum level and assessed the interactions between the lectin complement pathway (LCP) recognition molecules and MAP-1. We expressed recombinant MAP-1 in CHO DG44 cells, developed a quantitative ELISA assay based on a MAP-1 specific monoclonal capture antibody and measured the serum levels in 100 Danish blood donors. In addition we assessed the association properties between MAP-1 and Ficolin-2, -3 and MBL in serum using ELISA and density gradient ultra centrifugation. When recombinant MAP-1 was subjected to N-glycosidase F treatment the molecular mass decreased from ∼45. kDa to ∼40. kDa equivalent with the calculated molecular mass from the deduced amino acid sequence without the signal peptide. We found that serum MAP-1 was very stable when subjected to repeated freeze and thaw cycles. The mean serum concentration of MAP-1 was found to be 240. ng/ml (range: 115-466. ng/ml). MAP-1 was predominantly found in complex with Ficolin-3 and to a lesser degree with Ficolin-2 and MBL and by use of density gradient ultra centrifugation we could show that the major part of serum MAP-1 circulates in complex with the LCP molecules. In conclusion, these results show that MAP-1 is a highly stable glycosylated human serum protein found in complex with Ficolin-3, Ficolin-2 and MBL.

Original languageEnglish
JournalImmunobiology
Volume216
Issue number5
Pages (from-to)625-632
ISSN0171-2985
DOIs
Publication statusPublished - 1 May 2011

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