HEPARIN-BINDING LECTIN FROM HUMAN PLACENTA - FURTHER CHARACTERIZATION OF LIGAND-BINDING AND STRUCTURAL-PROPERTIES AND ITS RELATIONSHIP TO HISTONES AND HEPARIN-BINDING GROWTH-FACTORS

被引:41
作者
KOHNKEGODT, B [1 ]
GABIUS, HJ [1 ]
机构
[1] MAX PLANCK INST EXPTL MED,CHEM ABT,HERMANN REIN STR 3,W-3400 GOTTINGEN,GERMANY
关键词
D O I
10.1021/bi00215a009
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have previously demonstrated that the heparin-binding lectin of human placenta dissociates into up to four distinct polypeptides with molecular weights of 14 400, 15 000, 16 200, and 16 700 (Kohnke-Godt, B., & Gabius, H.-H. (1989) Biochemistry 28, 6531-6538). Stable complexes to ligands can shift the molecular weight appearance of the lectin to higher values. They can be dissociated in the additional presence of 9 M urea or by enzymatic degradation of heparin in model studies. The binding of heparin is rather stable over a range of salt concentrations from 1 to 3 M NaCl. Chemical modification with group-specific reagents to arginine, lysine, histidine, tyrosine, and tryptophan results in substantial inactivation of binding activity. Further amino-terminal sequence analyses point to a high-scoring relationship in this region to histone sequences, namely, histone H2B, but to no published sequences for any heparin-binding growth factor. Calculation of relatedness on the basis of differences in amino acid composition corroborates the conclusion of molecular distinction between the lectin, histones H2A and H2B, and the fibroblast growth factor as well as angiogenin. Histones only weakly agglutinate type II erythrocytes in contrast to the lectin. The immobilized lectin exhibits two classes of binding sites with K(D) values of 3 and 110 nM in contrast to one estimated K(D) value of 250 nM with a commercially available histone fraction. Both fractions retain binding activity to biotinylated heparin in transblots and are immunologically cross-reactive to antibodies, raised against the lectin as antigen. Subcellular fractionation clearly demonstrates that heparin-inhibitable hemagglutination activity and immunologically cross-reactive protein bands, characteristic for the lectin, but not unequivocally distinguishable from certain histone fractions in blots, are not confined to the nuclear fraction in the human placenta.
引用
收藏
页码:55 / 65
页数:11
相关论文
共 86 条
[1]  
ATEN RF, 1989, J BIOL CHEM, V264, P11065
[2]   MAST-CELL HEPARIN STIMULATES MIGRATION OF CAPILLARY ENDOTHELIAL-CELLS INVITRO [J].
AZIZKHAN, RG ;
AZIZKHAN, JC ;
ZETTER, BR ;
FOLKMAN, J .
JOURNAL OF EXPERIMENTAL MEDICINE, 1980, 152 (04) :931-944
[3]   EXPRESSION OF ENDOGENOUS RECEPTORS FOR NEOGLYCOPROTEINS, ESPECIALLY LECTINS, THAT ALLOW FIBER TYPING ON FORMALDEHYDE-FIXED, PARAFFIN-EMBEDDED MUSCLE BIOPSY SPECIMENS - A GLYCOHISTOCHEMICAL, IMMUNOHISTOCHEMICAL, AND GLYCOBIOCHEMICAL STUDY [J].
BARDOSI, A ;
DIMITRI, T ;
WOSGIEN, B ;
GABIUS, HJ .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1989, 37 (07) :989-998
[4]  
BARONDES SH, 1986, LECTINS PROPERTIES F, P437
[5]   DNA-BINDING TO HUMAN-LEUKOCYTES - EVIDENCE FOR A RECEPTOR-MEDIATED ASSOCIATION, INTERNALIZATION, AND DEGRADATION OF DNA [J].
BENNETT, RM ;
GABOR, GT ;
MERRITT, MM .
JOURNAL OF CLINICAL INVESTIGATION, 1985, 76 (06) :2182-2190
[6]   BINDING OF HEPARIN TO HUMAN MICROVASCULAR ENDOTHELIAL-CELLS AND THE EFFECT ON PROLIFERATION [J].
BIKFALVI, A ;
DUPUY, E ;
RUAN, C ;
TOBELEM, G ;
LESECHE, G ;
CAEN, J .
CELL BIOLOGY INTERNATIONAL REPORTS, 1988, 12 (11) :931-942
[7]   HEPARIN AND HEPARAN-SULFATE BINDING-SITES ON B-16 MELANOMA-CELLS [J].
BISWAS, C .
JOURNAL OF CELLULAR PHYSIOLOGY, 1988, 136 (01) :147-153
[8]  
BJORK I, 1989, BIOCHEMISTRY-US, V28, P1213
[9]   IMPROVED SILVER STAINING OF PLANT-PROTEINS, RNA AND DNA IN POLYACRYLAMIDE GELS [J].
BLUM, H ;
BEIER, H ;
GROSS, HJ .
ELECTROPHORESIS, 1987, 8 (02) :93-99
[10]   ISOLATION AND PARTIAL MOLECULAR CHARACTERIZATION OF PITUITARY FIBROBLAST GROWTH-FACTOR [J].
BOHLEN, P ;
BAIRD, A ;
ESCH, F ;
LING, N ;
GOSPODAROWICZ, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (17) :5364-5368