PMID: 8605233Feb 9, 1996Paper

Purification and characterization of N-acetylmuramyl-L-alanine amidase from human plasma using monoclonal antibodies

Biochimica Et Biophysica Acta
M A HoijerM P Hazenberg

Abstract

N-Acetylmuramyl-L-alanine amidase (EC 3.5.1.28) cleaves the amide bond between N-acetyl muramic acid and L-alanine in the peptide side chain of different peptidoglycan products. The enzyme was purified from human plasma using a three-step column chromatography procedure. Monoclonal antibodies were produced against the purified human enzyme. By coupling of a high affinity monoclonal antibody to sepharose beads an immunoadsorbent column was prepared. Using this second purification method it was possible to purify large amounts of the amidase from human plasma in a single step. SDS-PAGE showed one single band of 70 kDa and two-dimensional electrophoresis showed the presence of multiple isomeric forms of the protein with pI between 6.5 and 7.9. Two different methods were used for determination of substrate specificity, a HPLC method separating peptidoglycan monomers from the reaction products after incubation with amidase and a colorimetric method when high molecular weight peptidoglycan was used as a substrate for amidase. It is shown that the disaccharide tetra peptide, disaccharide penta peptide and the anhydro disaccharide tetrapeptide are good substrates for the amidase and that muramyl dipeptide and disaccharide dipeptide are...Continue Reading

References

Dec 1, 1977·The Journal of Experimental Medicine·W J CromartieC H Yang
Jul 6, 1990·Biochimica Et Biophysica Acta·E VanderwinkelJ P ten Have
Oct 1, 1985·Infection and Immunity·J Harrison, A Fox
Nov 18, 1981·Biochimica Et Biophysica Acta·B LadesićI Hrsak
Sep 1, 1984·Molecular and Cellular Biochemistry·P Jollès, J Jollès
May 1, 1995·Infection and Immunity·M A HoijerM P Hazenberg
Mar 1, 1993·American Journal of Clinical Pathology·M T SyrjäläT Krusius
Mar 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·K E LukerW E Goldman

❮ Previous
Next ❯

Citations

Dec 31, 2003·Molecular Immunology·Roman Dziarski
Jun 25, 2003·Biochemical and Biophysical Research Communications·Eva GeliusHåkan Steiner
Dec 22, 2015·Cellular & Molecular Immunology·Zhou ZhouBin Gao
Jun 18, 2004·Immunological Reviews·Håkan Steiner
Aug 2, 2005·Biochimica Et Biophysica Acta·Yinong ZhangRoman Dziarski
Aug 19, 2009·Biomedical Chromatography : BMC·Sze Kwan Lam, Tzi Bun Ng
Mar 4, 2006·Preparative Biochemistry & Biotechnology·Marina KrstanovićFabio Benedetti
Jul 26, 2017·Journal of Proteome Research·Melinda RezeliGyörgy Marko-Varga
Mar 17, 2020·International Journal of Toxicology·Kenneth L HastingsGary R Burleson
Sep 25, 2003·The Journal of Biological Chemistry·Zheng-Ming WangRoman Dziarski
Aug 26, 2017·Current Genetics·Roman Dziarski, Dipika Gupta
Apr 2, 2005·The Journal of Biological Chemistry·Christine McDonaldGabriel Nuñez
Jan 7, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Lizette VisserJon D Laman

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.