TY - JOUR
T1 - Fibulin-1C, C1 Esterase Inhibitor and Glucose Regulated Protein 75 Interact with the CREC Proteins, Calumenin and Reticulocalbin
AU - Hansen, G. A. W.
AU - Ludvigsen, M.
AU - Jacobsen, C.
AU - Cangemi, C.
AU - Rasmussen, L. M.
AU - Vorum, H.
AU - Honore, B.
N1 - Danish Medical Research Council [52-00-1004, 22-01-0249]; Novo Nordic Foundation; Danish Cancer Foundation [DS 02 071]; Danish Heart Foundation [08-4-R64-A2008-B894-22450] 1 e0132283 26161649
PY - 2015/7/10
Y1 - 2015/7/10
N2 - Affinity purification, immunoprecipitation, gel electrophoresis and mass spectrometry were used to identify fibulin-1C, C1 esterase inhibitor and glucose regulated protein 75, grp75, as binding partners of the CREC proteins, calumenin and reticulocalbin. Surface plasmon resonance was used to verify the interaction of all three proteins with each of the CREC proteins. Fibulin-1C interacts with calumenin and reticulocalbin with an estimated dissociation constant around 50-60 nM. The interaction, at least for reticulocalbin, was not dependent upon the presence of Ca
2+. C1 esterase inhibitor interacted with both proteins with an estimated dissociation constant at 1 μM for reticulocalbin and 150 nM for calumenin. The interaction, at least for calumenin, was dependent upon the presence of Ca
2+ with strong interaction at 3.5 mM while no detectable interaction could be found at 0.1 mM. Grp75 binds with an affinity of approximately 3-7 nM with reticulocalbin as well as with calumenin. These interactions suggest functional participation of the CREC proteins in chaperone activity, cell proliferation and transformation, cellular aging, haemostasis and thrombosis as well as modulation of the complement system in fighting bacterial infection.
AB - Affinity purification, immunoprecipitation, gel electrophoresis and mass spectrometry were used to identify fibulin-1C, C1 esterase inhibitor and glucose regulated protein 75, grp75, as binding partners of the CREC proteins, calumenin and reticulocalbin. Surface plasmon resonance was used to verify the interaction of all three proteins with each of the CREC proteins. Fibulin-1C interacts with calumenin and reticulocalbin with an estimated dissociation constant around 50-60 nM. The interaction, at least for reticulocalbin, was not dependent upon the presence of Ca
2+. C1 esterase inhibitor interacted with both proteins with an estimated dissociation constant at 1 μM for reticulocalbin and 150 nM for calumenin. The interaction, at least for calumenin, was dependent upon the presence of Ca
2+ with strong interaction at 3.5 mM while no detectable interaction could be found at 0.1 mM. Grp75 binds with an affinity of approximately 3-7 nM with reticulocalbin as well as with calumenin. These interactions suggest functional participation of the CREC proteins in chaperone activity, cell proliferation and transformation, cellular aging, haemostasis and thrombosis as well as modulation of the complement system in fighting bacterial infection.
KW - CALCIUM-BINDING PROTEIN MULTIPLE EF-HAND ENDOPLASMIC-RETICULUM CA2+-BINDING PROTEIN EXTRACELLULAR CALUMENIN PROTEOMIC APPROACH SECRETORY PATHWAY POTENTIAL ROLE CANCER CELLS EXPRESSION
KW - Amino Acid Sequence
KW - Arteries/metabolism
KW - Calcium-Binding Proteins/metabolism
KW - Cell Line
KW - Chromatography, Affinity
KW - Complement C1 Inhibitor Protein/metabolism
KW - Female
KW - HSP70 Heat-Shock Proteins/metabolism
KW - Humans
KW - Immune Sera/metabolism
KW - Immunohistochemistry
KW - Immunoprecipitation
KW - Mass Spectrometry
KW - Membrane Proteins/metabolism
KW - Microscopy, Confocal
KW - Molecular Sequence Data
KW - Peptides/chemistry
KW - Placenta/metabolism
KW - Pregnancy
KW - Protein Binding
KW - Reproducibility of Results
KW - Surface Plasmon Resonance
U2 - 10.1371/journal.pone.0132283
DO - 10.1371/journal.pone.0132283
M3 - Journal article
C2 - 26161649
VL - 10
JO - P L o S One
JF - P L o S One
SN - 1932-6203
IS - 7
M1 - e0132283
ER -