Structure and function of the N-linked glycans of HBP/CAP37/azurocidin: crystal structure determination and biological characterization of nonglycosylated HBP
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Structure and function of the N-linked glycans of HBP/CAP37/azurocidin : crystal structure determination and biological characterization of nonglycosylated HBP. / Iversen, L F; Kastrup, Jette Sandholm Jensen; Bjørn, S E; Wiberg, F C; Larsen, Ingrid K.; Flodgaard, H J; Rasmussen, P B.
In: Protein Science, Vol. 8, No. 10, 10.1999, p. 2019-26.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Structure and function of the N-linked glycans of HBP/CAP37/azurocidin
T2 - crystal structure determination and biological characterization of nonglycosylated HBP
AU - Iversen, L F
AU - Kastrup, Jette Sandholm Jensen
AU - Bjørn, S E
AU - Wiberg, F C
AU - Larsen, Ingrid K.
AU - Flodgaard, H J
AU - Rasmussen, P B
PY - 1999/10
Y1 - 1999/10
N2 - The three N-glycosylation sites of human heparin binding protein (HBP) have been mutated to produce a nonglycosylated HBP (ng-HBP) mutant. ng-HBP has been crystallized and tested for biological activity. Complete X-ray data have been collected to 2.1 A resolution, and the structure has been fully refined to an R-factor of 18.4% (R(free) 27.7%). The ng-HBP structure reveals that neither the secondary nor tertiary structure have changed due to the removal of the glycosylation, as compared to the previously determined glycosylated HBP structure. Although the primary events in N-linked glycosylation occurs concomitant with polypeptide synthesis and therefore possesses the ability to influence early events in protein folding, we see no evidence of glycosylation influencing the structure of the protein. The root-mean-square deviation between the superimposed structures was 0.24 A (on C alpha atoms), and only minor local structural differences are observed. Also, the overall stability of the protein seems to be unaffected by glycosylation, as judged by the B-factors derived from the two X-ray structures. The flexibility of a glycan site may be determined by the local polypeptide sequence and structure rather than the glycan itself. The biological in vitro activity assay data show that ng-HBP, contrary to glycosylated HBP, mediates only a very limited stimulation of the lipopolysaccharide induced cytokine release from human monocytes. In animal models of fecal peritonitis, glycosylated HBP treatment rescues mice from and an otherwise lethal injury. It appears that ng-HBP have significant effect on survival, and it can be concluded that ng-HBP can stimulate the host defence machinery albeit to a lesser extent than glycosylated HBP.
AB - The three N-glycosylation sites of human heparin binding protein (HBP) have been mutated to produce a nonglycosylated HBP (ng-HBP) mutant. ng-HBP has been crystallized and tested for biological activity. Complete X-ray data have been collected to 2.1 A resolution, and the structure has been fully refined to an R-factor of 18.4% (R(free) 27.7%). The ng-HBP structure reveals that neither the secondary nor tertiary structure have changed due to the removal of the glycosylation, as compared to the previously determined glycosylated HBP structure. Although the primary events in N-linked glycosylation occurs concomitant with polypeptide synthesis and therefore possesses the ability to influence early events in protein folding, we see no evidence of glycosylation influencing the structure of the protein. The root-mean-square deviation between the superimposed structures was 0.24 A (on C alpha atoms), and only minor local structural differences are observed. Also, the overall stability of the protein seems to be unaffected by glycosylation, as judged by the B-factors derived from the two X-ray structures. The flexibility of a glycan site may be determined by the local polypeptide sequence and structure rather than the glycan itself. The biological in vitro activity assay data show that ng-HBP, contrary to glycosylated HBP, mediates only a very limited stimulation of the lipopolysaccharide induced cytokine release from human monocytes. In animal models of fecal peritonitis, glycosylated HBP treatment rescues mice from and an otherwise lethal injury. It appears that ng-HBP have significant effect on survival, and it can be concluded that ng-HBP can stimulate the host defence machinery albeit to a lesser extent than glycosylated HBP.
KW - Animals
KW - Antimicrobial Cationic Peptides
KW - Base Sequence
KW - Blood Proteins
KW - Carrier Proteins
KW - Crystallography, X-Ray
KW - DNA Primers
KW - Glycosylation
KW - Humans
KW - Mice
KW - Polysaccharides
KW - Protein Conformation
KW - Recombinant Proteins
U2 - 10.1110/ps.8.10.2019
DO - 10.1110/ps.8.10.2019
M3 - Journal article
C2 - 10548047
VL - 8
SP - 2019
EP - 2026
JO - Protein Science
JF - Protein Science
SN - 0961-8368
IS - 10
ER -
ID: 44729507