Structure activity studies of the cytokine macrophage migration inhibitory factor (MIF) reveal a critical role for its carboxy terminus

R Mischke; A Gessner; A Kapurniotu; S Jüttner; R Kleemann; H Brunner; J Bernhagen

doi:10.1016/s0014-5793(97)01039-9

Structure activity studies of the cytokine macrophage migration inhibitory factor (MIF) reveal a critical role for its carboxy terminus

FEBS Lett. 1997 Sep 8;414(2):226-32. doi: 10.1016/s0014-5793(97)01039-9.

Authors

R Mischke¹, A Gessner, A Kapurniotu, S Jüttner, R Kleemann, H Brunner, J Bernhagen

Affiliation

¹ Laboratory of Biochemistry, Chair for Interfacial Engineering, University of Stuttgart, Germany.

PMID: 9315691
DOI: 10.1016/s0014-5793(97)01039-9

Abstract

Carboxy-truncated mutants of human MIF (MIF(1-104) and MIF(1-109)) were used in structure activity studies. CD spectroscopy revealed an overall structural similarity between the mutants and MIF. Denaturant-induced unfolding demonstrated that the C-terminus contributed significantly to the conformational stability of MIF. This appears to be due to the formation of two C-terminal beta-strands. The mutants were enzymatically active, exhibiting half of the enzymatic redox activity of MIF. However, immunological analysis showed that deletion of both 5 and 10 C-terminal residues resulted in loss of the macrophage activating properties of MIF, providing functional evidence that the C-terminus is important for immunological activity and trimer formation. A more detailed study of the C-terminus may assist in identifying the molecular basis for the immunological and enzymatic activities of MIF.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Antiprotozoal Agents
Biological Assay
Circular Dichroism
DNA Primers
Guanidine
Guanidines
Humans
Jurkat Cells
Leishmania major / drug effects
Macromolecular Substances
Macrophage Migration-Inhibitory Factors / biosynthesis
Macrophage Migration-Inhibitory Factors / chemistry*
Macrophage Migration-Inhibitory Factors / pharmacology*
Models, Structural
Molecular Sequence Data
Mutagenesis, Site-Directed
NADP Transhydrogenases / metabolism
Peptide Fragments / biosynthesis
Peptide Fragments / chemistry
Peptide Fragments / pharmacology
Polymerase Chain Reaction
Protein Conformation*
Protein Denaturation
Protein Structure, Secondary*
Recombinant Proteins / biosynthesis
Recombinant Proteins / chemistry
Recombinant Proteins / pharmacology
Structure-Activity Relationship
Thermodynamics

Substances

Antiprotozoal Agents
DNA Primers
Guanidines
Macromolecular Substances
Macrophage Migration-Inhibitory Factors
Peptide Fragments
Recombinant Proteins
NADP Transhydrogenases
Guanidine