Effect of enzymatic desialylation of human serum amyloid P component on surface exposure of laser photo CIDNP (chemically induced dynamic nuclear polarization)--reactive histidine, tryptophan and tyrosine residues.
The human pentraxin serum amyloid P component (SAP) exhibits no microheterogeneity in its complex di-antennary glycan. To elucidate whether the removal of sialic acids from this glycoprotein might affect the accessibility of certain amino acid residues of the protein we employed the laser photo CIDNP approach as a sensitive tool. The CIDNP effect is created by the interaction of a photoexcited dye with reactive amino acids and results in enhanced absorption- or emission-signals which can be observed for the three aromatic amino acids histidine, tryptophan, and tyrosine if they are accessible to the dye. Therefore, this technique can be applied to explore surface exposure of these residues. The respective spectra of SAP and enzymatically desialylated SAP were determined. Six tryptophan/histidine signals and one tyrosine signal are present in the aromatic part of the CIDNP difference spectrum of SAP. The corresponding spectrum of desialylated SAP shows remarkable alterations. The chemical shift of one tryptophan/histidine-characteristic signal is decreased by 0.1 ppm. One tryptophan/histidine signal disappeared and a new one was formed in the CIDNP difference spectrum of desialylated SAP, while the other signals were unaffected. The tyrosine signal has a clearly enhanced intensity in desialylated SAP. Therefore, the removal of sialic acid moieties from the single N-glycan of each monomer apparently affects surface presentation of distinct CIDNP-reactive amino acids of SAP.
Siebert
H C
HC
Institut für Physiologische Chemie, Tierärztliche Fakultät, Ludwig-Maximilians-Universität, München, Germany.
André
S
S
Reuter
G
G
Gabius
H J
HJ
Kaptein
R
R
Vliegenthart
J F
JF
D000596:Amino Acids / Q000032:analysis*; D005583:Fourier Analysis; D006639:Histidine / Q000032:analysis; D006801:Humans; D007834:Lasers; D009439:Neuraminidase; D010777:Photochemistry; D000683:Serum Amyloid P-Component / Q000737:chemistry*; D012794:Sialic Acids / Q000378:metabolism*; D013057:Spectrum Analysis / Q000379:methods*; D014364:Tryptophan / Q000032:analysis; D014443:Tyrosine / Q000032:analysis
D000596:Amino Acids; D000683:Serum Amyloid P-Component; D012794:Sialic Acids; D014443:Tyrosine; D006639:Histidine; D014364:Tryptophan; D009439:Neuraminidase