scholarly journals Analysis and identification of the Grem2 heparin/heparan sulfate-binding motif

2017 ◽  
Vol 474 (7) ◽  
pp. 1093-1107 ◽  
Author(s):  
Chandramohan Kattamuri ◽  
Kristof Nolan ◽  
Thomas B. Thompson
Keyword(s):  
Heparan Sulfate ◽  
Bone Morphogenetic Proteins ◽  
Family Members ◽  
Binding Motif ◽  
Heparin Binding ◽  
Surface Binding ◽  
Cell Surface Binding ◽  
Binding Epitope ◽  
Lysine Residues ◽  
The Individual

Bone morphogenetic proteins (BMPs) are regulated by extracellular antagonists of the DAN (differential screening-selected gene aberrative in neuroblastoma) family. Similar to the BMP ligands, certain DAN family members have been shown to interact with heparin and heparan sulfate (HS). Structural studies of DAN family members Gremlin-1 and Gremlin-2 (Grem2) have revealed a dimeric growth factor-like fold where a series of lysine residues cluster along one face of the protein. In the present study, we used mutagenesis, heparin-binding measurements, and cell surface-binding analysis to identify lysine residues that are important for heparin/HS binding in Grem2. We determined that residues involved in heparin/HS binding, while not necessary for BMP antagonism, merge with the heparin/HS-binding epitope of BMP2. Furthermore, the Grem2–BMP2 complex has higher affinity for heparin than the individual proteins and this affinity is not abrogated when the heparin/HS-binding epitope of Grem2 is attenuated. Overall, the present study shows that the Grem2 heparin/HS and BMP-binding epitopes are unique and independent, where, interestingly, the Grem2–BMP2 complex exhibits a significant increase in binding affinity toward heparin moieties that appear to be partially independent of the Grem2 heparin/HS-binding epitope.

scholarly journals Heparin and Activin-Binding Determinants in Follistatin and FSTL3

Endocrinology ◽  
2005 ◽  
Vol 146 (1) ◽  
pp. 130-136 ◽  
Author(s):  
Yisrael Sidis ◽  
Alan L. Schneyer ◽  
Henry T. Keutmann
Keyword(s):  
Cell Surface ◽  
Pituitary Cell ◽  
Heparin Binding ◽  
Surface Binding ◽  
Local Regulation ◽  
Cellular Processes ◽  
Cell Surface Binding ◽  
Binding Sequence ◽  
Binding Determinants ◽  
Heparin Affinity

Local regulation of pituitary FSH secretion and many other cellular processes by follistatin (FS) can be ascribed to its potent ability to bind and bioneutralize activin, in conjunction with binding to cell surface heparan-sulfate proteoglycans through a basic heparin-binding sequence (HBS; residues 75–86) in the first of the three FS domains. The FS homolog, FSTL3, also binds activin, but lacks any HBS and cannot associate with cell surfaces. We have used mutational analyses to define the determinants for heparin binding and activin interaction in FS and to determine the effects of conferring heparin binding to FSTL3. Mutants expressed from 283F cells were tested for cell surface and heparin affinity binding, for competititive activin binding and for bioactivity by suppression of pituitary cell FSH secretion. Replacement of the HBS or the full-length FS-domain 1 abolished cell surface binding but enhanced activin binding 4- to 8-fold. Surface binding was partially reduced after mutation of either lysine pair 75/76 or 81/82 and eliminated after mutation of both pairs. The 75/76 mutation reduced activin binding and, therefore, pituitary cell bioactivity by 5-fold. However, insertion of the HBS into FSTL3 did not restore heparin binding or pituitary-cell bioactivity. These results show that 1) the residues within the HBS are necessary but not sufficient for heparin binding, and 2) the HBS also harbors determinants for activin binding. Introduction of the full domain from FS conferred heparin binding to FSTL3, but activin binding was abolished. This implies an evolutionary safeguard against surface binding by FSTL3, supporting other evidence for physiological differences between FS and FSTL3.

Affinity of the heparin binding motif of Noggin1 to heparan sulfate and its visualization in the embryonic tissues

2015 ◽  
Vol 468 (1-2) ◽  
pp. 331-336 ◽  
Author(s):  
Alexey M. Nesterenko ◽  
Eugeny E. Orlov ◽  
Galina V. Ermakova ◽  
Igor A. Ivanov ◽  
Pavel I. Semenyuk ◽  
...  
Keyword(s):  
Heparan Sulfate ◽  
Binding Motif ◽  
Heparin Binding ◽  
Embryonic Tissues

Ephrin-B3 binds to a sulfated cell-surface receptor

2010 ◽  
Vol 433 (1) ◽  
pp. 215-223 ◽  
Author(s):  
Halvor L. Holen ◽  
Lillian Zernichow ◽  
Kristine E. Fjelland ◽  
Ida M. Evenroed ◽  
Kristian Prydz ◽  
...  
Keyword(s):  
Cell Surface ◽  
Heparan Sulfate ◽  
Simian Virus 40 ◽  
T Antigen ◽  
Site Directed Mutagenesis ◽  
Cell Surface Receptor ◽  
Binding Property ◽  
Heparin Binding ◽  
Surface Binding ◽  
Cellular Signalling

The ephrins are a family of proteins known to bind the Eph (erythropoietin-producing hepatocellular) receptor tyrosine kinase family. In the present paper, we provide data showing that ephrin-B3 binds a sulfated cell-surface protein on HEK-293T (human embryonic kidney-293 cells expressing the large T-antigen of simian virus 40) and HeLa cells, a binding that is nearly completely blocked by treatment of these cell lines with chlorate or heparinase, or by addition of the heavily sulfated glycosaminoglycan heparin. This indicates that heparan sulfate on these cells is essential for cell-surface binding of ephrin-B3. Heparin did not affect ephrin-B3 binding to EphB receptors expressed on transfected HEK-293T cells, indicating further that ephrin-B3 binds an alternative receptor which is a heparan sulfate proteoglycan. Site-directed mutagenesis analysis revealed that Arg178 and Lys179 are important for heparin binding of ephrin-B3 and also for ephrin-B3 binding to cells. These amino acids, when introduced in the non-heparin-binding ephrin-B1, conferred the heparin-binding property. Functional studies reveal that ephrin-B3 binding to cells induces cellular signalling and influences cell rounding and cell spreading. In conclusion, our data provide evidence for an unknown ephrin-B3-binding cell-surface proteoglycan involved in cellular signalling.

scholarly journals Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns.

1994 ◽  
Vol 5 (7) ◽  
pp. 797-805 ◽  
Author(s):  
C W Kim ◽  
O A Goldberger ◽  
R L Gallo ◽  
M Bernfield
Keyword(s):  
Heparan Sulfate ◽  
Family Member ◽  
Cultured Cells ◽  
Family Members ◽  
Cell Types ◽  
Heparan Sulfate Proteoglycans ◽  
Mrna Levels ◽  
Cell Tissue ◽  
Mouse Tissues ◽  
The Individual

The syndecans are a gene family of four transmembrane heparan sulfate proteoglycans that bind, via their HS chains, diverse components of the cellular microenvironment. To evaluate the expression of the individual syndecans, we prepared cDNA probes to compare mRNA levels in various adult mouse tissues and cultured mouse cells representing various epithelial, fibroblastic, endothelial, and neural cell types and B cells at various stages of differentiation. We also prepared antibody probes to assess whether the extracellular domains of the individual syndecans are shed into the conditioned media of cultured cells. Our results show that all cells and tissues studied, except B-stem cells, express at least one syndecan family member; most cells and tissues express multiple syndecans. However, each syndecan family member is expressed selectively in cell-, tissue-, and development-specific patterns. The extracellular domain of all syndecan family members is shed as an intact proteoglycan. Thus, most, if not all, cells acquire a distinctive repertoire of the four syndecan family members as they differentiate, resulting in selective patterns of expression that likely reflect distinct functions.

scholarly journals Deamidation and glycation of a Bacillus licheniformis α-amylase during industrial fermentation can improve detergent wash performance

Amylase ◽  
2021 ◽  
Vol 5 (1) ◽  
pp. 38-49
Author(s):  
Connie Pontoppidan ◽  
Svend G. Kaasgaard ◽  
Carsten P. Sønksen ◽  
Carsten Andersen ◽  
Birte Svensson
Keyword(s):  
Bacillus Licheniformis ◽  
Mutational Analysis ◽  
Peptide Mapping ◽  
Substrate Binding ◽  
Carbohydrate Binding ◽  
Surface Binding ◽  
Lysine Residues ◽  
Tandem Mass Spectrometric ◽  
Binding Cleft ◽  
Household Detergents

Abstract The industrial thermostable Bacillus licheniformis α-amylase (BLA) has wide applications, including in household detergents, and efforts to improve its performance are continuously ongoing. BLA during the industrial production is deamidated and glycated resulting in multiple forms with different isoelectric points. Forty modified positions were identified by tandem mass spectrometric peptide mapping of BLA forms separated by isoelectric focusing. These modified 12 asparagine, 9 glutamine, 8 arginine and 11 lysine residues are mostly situated on the enzyme surface and several belong to regions involved in stability, activity and carbohydrate binding. Eight residues presumed to interact with starch at the active site and surface binding sites (SBSs) were subjected to mutational analysis. Five mutants mimicking deamidation (N→D, Q→E) at the substrate binding cleft showed moderate to no effect on thermostability and k cat and K M for maltoheptaose and amylose. Notably, the mutations improved laundry wash efficiency in detergents at pH 8.5 and 10.0. Replacing three reducing sugar reactive side chains (K→M, R→L) at a distant substrate binding region and two SBSs enhanced wash performance especially in liquid detergent at pH 8.5, slightly improved enzymatic activity and maintained thermostability. Wash performance was most improved (5-fold) for the N265D mutant near substrate binding subsite +3.

scholarly journals Identification of a novel cell attachment domain in the HIV-1 Tat protein and its 90-kDa cell surface binding protein.

1993 ◽  
Vol 268 (7) ◽  
pp. 5279-5284
Author(s):  
B.S. Weeks ◽  
K. Desai ◽  
P.M. Loewenstein ◽  
M.E. Klotman ◽  
P.E. Klotman ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Attachment ◽  
Binding Protein ◽  
Tat Protein ◽  
Surface Binding ◽  
Cell Surface Binding ◽  
Hiv 1

scholarly journals Cell surface-binding sites for progesterone mediate calcium uptake in human sperm.

1991 ◽  
Vol 266 (28) ◽  
pp. 18655-18659 ◽  
Author(s):  
P.F. Blackmore ◽  
J. Neulen ◽  
F. Lattanzio ◽  
S.J. Beebe
Keyword(s):  
Cell Surface ◽  
Binding Sites ◽  
Calcium Uptake ◽  
Human Sperm ◽  
Surface Binding ◽  
Cell Surface Binding
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