Subdomain

14,000,000 Leading Edge Experts on the ideXlab platform

Scan Science and Technology

Contact Leading Edge Experts & Companies

Scan Science and Technology

Contact Leading Edge Experts & Companies

The Experts below are selected from a list of 65340 Experts worldwide ranked by ideXlab platform

A Laine - One of the best experts on this subject based on the ideXlab platform.

  • Human mucin gene MUC5B, the 10.7-kb large central exon encodes various alternate Subdomains resulting in a super-repeat. Structural evidence for a 11p15.5 gene family.
    Journal of Biological Chemistry, 1997
    Co-Authors: Jean-luc Desseyn, V Guyonnet-dupérat, N Porchet, J P Aubert, A Laine
    Abstract:

    Human mucin gene MUC5B is mapped clustered with MUC6, MUC2, and MUC5AC on chromosome 11p15.5. We report here the isolation of three overlapping genomic clones of human MUC5B spanning approximately 40 kilobases. We have determined their partial restriction maps and the intron-exon boundaries of the central region encoding a single open reading frame. This coding region has been completely sequenced. Its length is 10,713 base pairs, and it encodes a 3570-amino acid peptide. Nineteen Subdomains have been individualized. Some Subdomains show similarity to each other, creating larger composite repeat units that we have called super-repeats. Four super-repeats of 528 amino acid residues are thus observed within the central exon. Each comprises (i) a Subdomain composed of 11 repeats of the irregular repeat of 29 amino acid residues, (ii) a unique conserved Subdomain with no typical repeat, and (iii) a cysteine-rich Subdomain. This latter Subdomain has high sequence similarity to the cysteine-rich domains described in MUC2 and MUC5AC. Sequence data of these three genes, together with their clustered organization, lead us to suggest that they may be a part of a multigene family. The super-repeat present in MUC5B is the largest ever determined in mucin genes and the central exon of this gene is, by far, the largest reported for a vertebrate gene.

  • human mucin gene muc5b the 10 7 kb large central exon encodes various alternate Subdomains resulting in a super repeat structural evidence for a 11p15 5 gene family
    Journal of Biological Chemistry, 1997
    Co-Authors: Jean-luc Desseyn, N Porchet, Veronique Guyonnetduperat, Jeanpierre Aubert, A Laine
    Abstract:

    Abstract Human mucin gene MUC5B is mapped clustered with MUC6, MUC2, and MUC5AC on chromosome 11p15.5. We report here the isolation of three overlapping genomic clones of human MUC5B spanning approximately 40 kilobases. We have determined their partial restriction maps and the intron-exon boundaries of the central region encoding a single open reading frame. This coding region has been completely sequenced. Its length is 10,713 base pairs, and it encodes a 3570-amino acid peptide. Nineteen Subdomains have been individualized. Some Subdomains show similarity to each other, creating larger composite repeat units that we have called super-repeats. Four super-repeats of 528 amino acid residues are thus observed within the central exon. Each comprises (i) a Subdomain composed of 11 repeats of the irregular repeat of 29 amino acid residues, (ii) a unique conserved Subdomain with no typical repeat, and (iii) a cysteine-rich Subdomain. This latter Subdomain has high sequence similarity to the cysteine-rich domains described in MUC2 and MUC5AC. Sequence data of these three genes, together with their clustered organization, lead us to suggest that they may be a part of a multigene family. The super-repeat present in MUC5B is the largest ever determined in mucin genes and the central exon of this gene is, by far, the largest reported for a vertebrate gene.

Jan Bogerd - One of the best experts on this subject based on the ideXlab platform.

  • ligand selectivity of gonadotropin receptors role of the β strands of extracellular leucine rich repeats 3 and 6 of the human luteinizing hormone receptor
    Journal of Biological Chemistry, 2003
    Co-Authors: Henry F. Vischer, Joke C. M. Granneman, Michiel J. Noordam, Sietse Mosselman, Jan Bogerd
    Abstract:

    The difference in hormone selectivity between the human follicle-stimulating hormone receptor (hFSH-R) and human luteinizing hormone/chorionic gonadotropin receptor (hLH-R) is determined by their approximately 350 amino acid-long N-terminal receptor exodomains that allow the mutually exclusive binding of human follicle-stimulating hormone (hFSH) and human luteinizing hormone (hLH) when these hormones are present in physiological concentrations. The exodomains of each of these receptors consist of a nine-leucine-rich repeat-containing Subdomain (LRR Subdomain) flanked by N- and C-terminal cysteine-rich Subdomains. Chimeric receptors, in which the structural Subdomains of the hFSH-R exodomain were substituted with those of the hLH-R, showed a similar high responsiveness to human chorionic gonadotropin (hCG) and hLH as long as they harbored the LRR Subdomain of the hLH-R. In addition, these chimeric receptors showed no responsiveness to hFSH. The LRR Subdomains of the gonadotropin receptor exodomains are predicted to adopt a horseshoe-like conformation, of which the hormone-binding concave surface is composed of nine parallel beta-strands. Receptors in which individual beta-strands of the hFSH-R were replaced with the corresponding hLH-R sequences revealed that hCG and hLH selectivity is predominantly determined by hLH-R beta-strands 3 and 6. A mutant receptor in which the hFSH-R beta-strands 3 and 6 were substituted simultaneously with their hLH-R counterparts displayed a responsiveness to hCG and hLH similar to that of the wild type hLH-R. Responsiveness to hFSH was not affected by most beta-strand substitutions, suggesting the involvement of multiple low-impact determinants for this hormone.

  • Ligand selectivity of gonadotropin receptors. Role of the beta-strands of extracellular leucine-rich repeats 3 and 6 of the human luteinizing hormone receptor.
    The Journal of biological chemistry, 2003
    Co-Authors: Henry F. Vischer, Joke C. M. Granneman, Michiel J. Noordam, Sietse Mosselman, Jan Bogerd
    Abstract:

    The difference in hormone selectivity between the human follicle-stimulating hormone receptor (hFSH-R) and human luteinizing hormone/chorionic gonadotropin receptor (hLH-R) is determined by their approximately 350 amino acid-long N-terminal receptor exodomains that allow the mutually exclusive binding of human follicle-stimulating hormone (hFSH) and human luteinizing hormone (hLH) when these hormones are present in physiological concentrations. The exodomains of each of these receptors consist of a nine-leucine-rich repeat-containing Subdomain (LRR Subdomain) flanked by N- and C-terminal cysteine-rich Subdomains. Chimeric receptors, in which the structural Subdomains of the hFSH-R exodomain were substituted with those of the hLH-R, showed a similar high responsiveness to human chorionic gonadotropin (hCG) and hLH as long as they harbored the LRR Subdomain of the hLH-R. In addition, these chimeric receptors showed no responsiveness to hFSH. The LRR Subdomains of the gonadotropin receptor exodomains are predicted to adopt a horseshoe-like conformation, of which the hormone-binding concave surface is composed of nine parallel beta-strands. Receptors in which individual beta-strands of the hFSH-R were replaced with the corresponding hLH-R sequences revealed that hCG and hLH selectivity is predominantly determined by hLH-R beta-strands 3 and 6. A mutant receptor in which the hFSH-R beta-strands 3 and 6 were substituted simultaneously with their hLH-R counterparts displayed a responsiveness to hCG and hLH similar to that of the wild type hLH-R. Responsiveness to hFSH was not affected by most beta-strand substitutions, suggesting the involvement of multiple low-impact determinants for this hormone.

Richard W. Kriwacki - One of the best experts on this subject based on the ideXlab platform.

  • incomplete folding upon binding mediates cdk4 cyclin d complex activation by tyrosine phosphorylation of inhibitor p27 protein
    Journal of Biological Chemistry, 2011
    Co-Authors: Antonio M. Ferreira, Steve Otieno, Limin Xiao, Donald Bashford, Richard W. Kriwacki
    Abstract:

    Abstract p27Kip1 (p27), an intrinsically disordered protein, regulates the various Cdk/cyclin complexes that control cell cycle progression. The kinase inhibitory domain of p27 contains a cyclin-binding Subdomain (D1), a Cdk-binding Subdomain (D2), and a linker helix Subdomain that connects D1 and D2. Here, we report that, despite extensive sequence conservation between Cdk4/cyclin D1 (hereafter Cdk4/cyclin D) and Cdk2/cyclin A, the thermodynamic details describing how the individual p27 Subdomains contribute to equally high affinity binding to these two Cdk/cyclin complexes are strikingly different. Differences in enthalpy/entropy compensation revealed that the D2 Subdomain of p27 folds incompletely when binding Cdk4/cyclin D versus Cdk2/cyclin A. Incomplete binding-induced folding exposes tyrosine 88 of p27 for phosphorylation by the nonreceptor tyrosine kinase Abl. Importantly, tyrosine phosphorylation (of p27) relieves Cdk inhibition by p27, enabling cell cycle entry. Furthermore, the interaction between a conserved hydrophobic patch on cyclin D and Subdomain D1 is much weaker than that with cyclin A; consequently, a construct containing Subdomains D1 and LH (p27-D1LH) does not inhibit substrate binding to Cdk4/cyclin D as it does to Cdk2/cyclin A. Our results provide a mechanism by which Cdk4 (within the p27/Cdk4/cyclin D complex) is poised to be activated by extrinsic mitogenic signals that impinge upon p27 at the earliest stage of cell division. More broadly, our results further illustrate the regulatory versatility of intrinsically disordered proteins.

  • Incomplete Folding upon Binding Mediates Cdk4/Cyclin D Complex Activation by Tyrosine Phosphorylation of Inhibitor p27 Protein
    Journal of Biological Chemistry, 2011
    Co-Authors: Antonio M. Ferreira, Steve Otieno, Limin Xiao, Donald Bashford, Richard W. Kriwacki
    Abstract:

    Abstract p27Kip1 (p27), an intrinsically disordered protein, regulates the various Cdk/cyclin complexes that control cell cycle progression. The kinase inhibitory domain of p27 contains a cyclin-binding Subdomain (D1), a Cdk-binding Subdomain (D2), and a linker helix Subdomain that connects D1 and D2. Here, we report that, despite extensive sequence conservation between Cdk4/cyclin D1 (hereafter Cdk4/cyclin D) and Cdk2/cyclin A, the thermodynamic details describing how the individual p27 Subdomains contribute to equally high affinity binding to these two Cdk/cyclin complexes are strikingly different. Differences in enthalpy/entropy compensation revealed that the D2 Subdomain of p27 folds incompletely when binding Cdk4/cyclin D versus Cdk2/cyclin A. Incomplete binding-induced folding exposes tyrosine 88 of p27 for phosphorylation by the nonreceptor tyrosine kinase Abl. Importantly, tyrosine phosphorylation (of p27) relieves Cdk inhibition by p27, enabling cell cycle entry. Furthermore, the interaction between a conserved hydrophobic patch on cyclin D and Subdomain D1 is much weaker than that with cyclin A; consequently, a construct containing Subdomains D1 and LH (p27-D1LH) does not inhibit substrate binding to Cdk4/cyclin D as it does to Cdk2/cyclin A. Our results provide a mechanism by which Cdk4 (within the p27/Cdk4/cyclin D complex) is poised to be activated by extrinsic mitogenic signals that impinge upon p27 at the earliest stage of cell division. More broadly, our results further illustrate the regulatory versatility of intrinsically disordered proteins.

  • The Role of the LH Subdomain in the Function of the Cip/Kip Cyclin-Dependent Kinase Regulators
    Biophysical journal, 2011
    Co-Authors: Steve Otieno, Christy R. Grace, Richard W. Kriwacki
    Abstract:

    The Cip/Kip protein family, which includes p27, p21, and p57, modulates the activity of cyclin-dependent kinases (Cdks). A domain within these proteins, termed the kinase inhibitory domain (KID), is necessary and sufficient for Cdk inhibition. The KID consists of a cyclin-binding Subdomain (termed D1) and a Cdk-binding Subdomain (termed D2) joined by a 22-residue linker Subdomain (termed LH). Before binding the Cdks, D1 and D2 are largely unstructured and the LH Subdomain exhibits nascent helical characteristics. Curiously, although the sequence of the linker Subdomain is not highly conserved within the family, its nascent helical structure is conserved. In this study, we explored the role of this structural conservation in interactions with cyclin-dependent kinase 2 (Cdk2) and cyclin A. We constructed chimeric p27-KID molecules in which the p27 LH Subdomain was replaced with the corresponding segments of either p21 or p57. The chimeric molecules bind and inhibit Cdk2 in a manner similar to wild-type p27-KID. However, the extent of enthalpy/entropy compensation associated with these interactions was dramatically different, indicating different extents of LH Subdomain folding upon binding. Our results indicate that the different LH Subdomains, despite their sequence and thermodynamic differences, play similar roles in binding and inhibiting Cdk2/cyclin A.

Jean-luc Desseyn - One of the best experts on this subject based on the ideXlab platform.

  • Human mucin gene MUC5B, the 10.7-kb large central exon encodes various alternate Subdomains resulting in a super-repeat. Structural evidence for a 11p15.5 gene family.
    Journal of Biological Chemistry, 1997
    Co-Authors: Jean-luc Desseyn, V Guyonnet-dupérat, N Porchet, J P Aubert, A Laine
    Abstract:

    Human mucin gene MUC5B is mapped clustered with MUC6, MUC2, and MUC5AC on chromosome 11p15.5. We report here the isolation of three overlapping genomic clones of human MUC5B spanning approximately 40 kilobases. We have determined their partial restriction maps and the intron-exon boundaries of the central region encoding a single open reading frame. This coding region has been completely sequenced. Its length is 10,713 base pairs, and it encodes a 3570-amino acid peptide. Nineteen Subdomains have been individualized. Some Subdomains show similarity to each other, creating larger composite repeat units that we have called super-repeats. Four super-repeats of 528 amino acid residues are thus observed within the central exon. Each comprises (i) a Subdomain composed of 11 repeats of the irregular repeat of 29 amino acid residues, (ii) a unique conserved Subdomain with no typical repeat, and (iii) a cysteine-rich Subdomain. This latter Subdomain has high sequence similarity to the cysteine-rich domains described in MUC2 and MUC5AC. Sequence data of these three genes, together with their clustered organization, lead us to suggest that they may be a part of a multigene family. The super-repeat present in MUC5B is the largest ever determined in mucin genes and the central exon of this gene is, by far, the largest reported for a vertebrate gene.

  • human mucin gene muc5b the 10 7 kb large central exon encodes various alternate Subdomains resulting in a super repeat structural evidence for a 11p15 5 gene family
    Journal of Biological Chemistry, 1997
    Co-Authors: Jean-luc Desseyn, N Porchet, Veronique Guyonnetduperat, Jeanpierre Aubert, A Laine
    Abstract:

    Abstract Human mucin gene MUC5B is mapped clustered with MUC6, MUC2, and MUC5AC on chromosome 11p15.5. We report here the isolation of three overlapping genomic clones of human MUC5B spanning approximately 40 kilobases. We have determined their partial restriction maps and the intron-exon boundaries of the central region encoding a single open reading frame. This coding region has been completely sequenced. Its length is 10,713 base pairs, and it encodes a 3570-amino acid peptide. Nineteen Subdomains have been individualized. Some Subdomains show similarity to each other, creating larger composite repeat units that we have called super-repeats. Four super-repeats of 528 amino acid residues are thus observed within the central exon. Each comprises (i) a Subdomain composed of 11 repeats of the irregular repeat of 29 amino acid residues, (ii) a unique conserved Subdomain with no typical repeat, and (iii) a cysteine-rich Subdomain. This latter Subdomain has high sequence similarity to the cysteine-rich domains described in MUC2 and MUC5AC. Sequence data of these three genes, together with their clustered organization, lead us to suggest that they may be a part of a multigene family. The super-repeat present in MUC5B is the largest ever determined in mucin genes and the central exon of this gene is, by far, the largest reported for a vertebrate gene.

Edward E Morrisey - One of the best experts on this subject based on the ideXlab platform.

  • transcriptional and dna binding activity of the foxp1 2 4 family is modulated by heterotypic and homotypic protein interactions
    Molecular and Cellular Biology, 2004
    Co-Authors: Shanru Li, Joel Weidenfeld, Edward E Morrisey
    Abstract:

    Foxp1, Foxp2, and Foxp4 are large multidomain transcriptional regulators belonging to the family of winged-helix DNA binding proteins known as the Fox family. Foxp1 and Foxp2 have been shown to act as transcriptional repressors, while regulatory activity of the recently identified Foxp4 has not been determined. Given the importance of this Fox gene subfamily in neural and lung development, we sought to elucidate the mechanisms by which Foxp1, Foxp2, and Foxp4 repress gene transcription. We show that like Foxp1 and Foxp2, Foxp4 represses transcription. Analysis of the N-terminal repression domain in Foxp1, Foxp2, and Foxp4 shows that this region contains two separate and distinct repression Subdomains that are highly homologous termed Subdomain 1 and Subdomain 2. However, Subdomain 2 is not functional in Foxp4. Screening for proteins that interact with Subdomains 1 and 2 of Foxp2 using yeast two-hybrid analysis revealed that Subdomain 2 binds to C-terminal binding protein 1, which can synergistically repress transcription with Foxp1 and Foxp2, but not Foxp4. Subdomain 1 contains a highly conserved leucine zipper similar to that found in N-myc and confers homo- and heterodimerization to the Foxp1/2/4 family members. These interactions are dependent on the conserved leucine zipper motif. Finally, we show that the integrity of this Subdomain is essential for DNA binding, making Foxp1, Foxp2, and Foxp4 the first Fox proteins that require dimerization for DNA binding. These data reveal a complex regulatory mechanism underlying Foxp1, Foxp2, and Foxp4 activity, demonstrating that Foxp1, Foxp2, and Foxp4 are the first Fox proteins reported whose activity is regulated by homo- and heterodimerization.

  • Transcriptional and DNA Binding Activity of the Foxp1/2/4 Family Is Modulated by Heterotypic and Homotypic Protein Interactions
    Molecular and cellular biology, 2004
    Co-Authors: Joel Weidenfeld, Edward E Morrisey
    Abstract:

    Foxp1, Foxp2, and Foxp4 are large multidomain transcriptional regulators belonging to the family of winged-helix DNA binding proteins known as the Fox family. Foxp1 and Foxp2 have been shown to act as transcriptional repressors, while regulatory activity of the recently identified Foxp4 has not been determined. Given the importance of this Fox gene subfamily in neural and lung development, we sought to elucidate the mechanisms by which Foxp1, Foxp2, and Foxp4 repress gene transcription. We show that like Foxp1 and Foxp2, Foxp4 represses transcription. Analysis of the N-terminal repression domain in Foxp1, Foxp2, and Foxp4 shows that this region contains two separate and distinct repression Subdomains that are highly homologous termed Subdomain 1 and Subdomain 2. However, Subdomain 2 is not functional in Foxp4. Screening for proteins that interact with Subdomains 1 and 2 of Foxp2 using yeast two-hybrid analysis revealed that Subdomain 2 binds to C-terminal binding protein 1, which can synergistically repress transcription with Foxp1 and Foxp2, but not Foxp4. Subdomain 1 contains a highly conserved leucine zipper similar to that found in N-myc and confers homo- and heterodimerization to the Foxp1/2/4 family members. These interactions are dependent on the conserved leucine zipper motif. Finally, we show that the integrity of this Subdomain is essential for DNA binding, making Foxp1, Foxp2, and Foxp4 the first Fox proteins that require dimerization for DNA binding. These data reveal a complex regulatory mechanism underlying Foxp1, Foxp2, and Foxp4 activity, demonstrating that Foxp1, Foxp2, and Foxp4 are the first Fox proteins reported whose activity is regulated by homo- and heterodimerization.