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Current Protein and Peptide Science, Vol. 2, No. 2, 2001

Contents

Fusion Proteins from Artificial and Natural Structural Modules Pp.107-121

N. Liu, G. Caderas, C. Deillon, S. Hoffmann, S. Klauser, T. Cui and B. Gutte

[Abstract]

 

Revisiting proteus: Do Minor Changes in Lectin Structure Matter in Biological Activity? Lessons from and Potential Biotechnological Uses of the Diocleinae Subtribe Lectins Pp.123-135

B. S. Cavada T. Barbosa, S. Arruda, T. B. Grangeiro and M. Barral-Netto

[Abstract]

 

Inhibition of Cysteine Protease Activity by NO-donors Pp.137-153

Paolo Ascenzi, Luca Salvati, Martino Bolognesi Marco Colasanti Fabio Polticelli and Giorgio Venturini

[Abstract]

 

How The Sequestration of a Protein Interferes with its Mechanism of Action: Example of a New Family of Proteins Characterized by a Particular Cysteine-Rich Carboxy-Terminal Domain Involved in Gene Expression Regulation Pp.155-167

S.1 Thébault and J.M. Mesnard

[Abstract]

 

Mif1: A Missing Link between the Unfolded Protein Response Pathway and ER-Associated Protein

Degradation? Pp.169-190

Theo van Laar, Alex J. van der Eb and Carrol Terleth

[Abstract]


Abstracts

 

[Back to top]  Fusion Proteins from Artificial and Natural Structural Modules

N. Liu, G. Caderas, C. Deillon, S. Hoffmann, S. Klauser, T. Cui and B. Gutte

 

The purpose of preparing fusion proteins from designed and natural sequences is mainly twofold; it aims at the stabilization of structure and at the modification of biological activity. Fusion with b-galactosidase, for example, can increase the intracellular stability and DDT-degrading activity of an artificial DDT-binding peptide, and fusions with a leucine zipper produce mono- and bifunctional single-chain variable domain antibody fragments or homodimeric and heterodimeric DNA-binding proteins like an artificial homodimeric HIV-1 enhancer-binding protein with increased binding specificity and repressor activity. Of importance are also short leader sequences that mediate the translocation of proteins across the cytoplasmic and the nuclear membrane. An interesting by-product of the leucine zipper-mediated dimerization of an HIV-1 enhancer-binding protein was the synthesis and the structural as well as functional characterization of a retro-leucine zipper.

 

[Back to top] Revisiting proteus: Do Minor Changes in Lectin Structure Matter in Biological Activity? Lessons from and Potential Biotechnological Uses of the Diocleinae Subtribe Lectins

B. S. Cavada T. Barbosa, S. Arruda, T. B. Grangeiro and M. Barral-Netto

 

Significant differences in function have been observed among lectins structurally similar to concanavalin A, but their high homology with this widely used lectin has kept them in obscurity. The observation of large differences in the potency of many of these Diocleinae lectins as stimulators of Interferon-g production by human peripheral blood mononuclear cells has lead to a major effort to unravel their chemical structure and biological activity. Modeling studies of some of these lectins reveal conformational changes in side chains of some residues involved in the carbohydrate-binding site, with possible effects on the ability of these proteins to recognize specific carbohydrate structures. Additionally, all them constitute in fact a mixture of isolectins, which in different proportions could lead to diverse effects. The present review of the biological actions of Diocleinae lectins includes several in vitro and in vivo immunological findings, as well as their effects on insect growth and reproduction. In these systems Diocleinae lectins proved to be quite diverse in their potency. Such diversity in the biological activity of highly related proteins recalls the origin of the name protein: like Proteus, the capability of assuming various forms is the essential feature of this class of molecules.

 

[Back to top] Inhibition of Cysteine Protease Activity by NO-donors

Paolo Ascenzi, Luca Salvati, Martino Bolognesi Marco Colasanti Fabio Polticelli and Giorgio Venturini

 

Cysteine proteases represent a broad class of proteolytic enzymes widely distributed among living organisms. Although well known as typical lysosomal enzymes, cysteine proteases are actually recognized as multi-function enzymes, being involved in antigen processing and presentation, in membrane-bound protein cleavage, as well as in degradation of the cellular matrix and in processes of tissue remodeling. Very recently, it has been shown that the NO(-donor)-mediated chemical modification of the Cys catalytic residue of cysteine proteases, including Coxsackievirus and Rhinovirus cysteine proteases, cruzain, Leishmania infantum cysteine protease, falcipain, papain, as well as mammalian caspases, cathepsins and calpain, blocks the enzyme activity in vitro and in vivo. Here, inhibition of representative cysteine proteases by NO(-donors) is reviewed.

 

[Back to top]  How The Sequestration of a Protein Interferes with its Mechanism of Action: Example of a New Family of Proteins Characterized by a Particular Cysteine-Rich Carboxy-Terminal Domain Involved in Gene Expression Regulation

S.1 Thébault and J.M. Mesnard

 

We describe here a new family of proteins characterized by a particular cysteine-rich carboxy-terminal domain and involved in gene expression regulation. This family presently includes three members: I-mfa (inhibitor of MyoD family), HIC p40 and HIC p32 (human I-mfa domain-containing protein). I-mfa, by interacting with MyoD family members, represses both transcriptional activation and myogenesis mediated by these factors. HIC two isoforms, HIC p40 and HIC p32, are involved in the positive regulation of Tax-mediated HTLV-I (human T-cell leukemia virus type 1) promoter activation and in the negative regulation of Tat-mediated HIV-1 (human immunodeficiency virus type 1) promoter transcription. The common carboxy-terminal region of HIC p40 and HIC p32, which is clearly involved in these regulations, shares 77% homology with the carboxy-terminal domain of I-mfa. This suggests that I-mfa, HIC p40 and HIC p32 are part of a new family of proteins involved in gene expression regulation and characterized by a specific cysteine-rich carboxy-terminal domain. Moreover, the three proteins present different subcellular localizations: I-mfa and HIC p32 are mainly cytoplasmic while HIC p40 is mainly nucleolar. The specific localization of each member of this new family will be discussed, possibly explaining how they work. Effectively, a mechanism of protein sequestration in a particular compartment, cytoplasm or nucleolus, could be involved in their function, as it is the case for many other proteins. This relationship between sequestration and function regulation will be exemplified for several cellular factors.

 

[Back to top] Mif1: A Missing Link between the Unfolded Protein Response Pathway and ER-Associated Protein Degradation?

Theo van Laar, Alex J. van der Eb and Carrol Terleth

 

Eukaryotic cells have three different mechanisms to deal with the accumulation of unfolded proteins in the endoplasmic reticulum: (1) In cells in which unfolded polypeptides accumulate, translation initiation is inhibited to prevent further accumulation of unfolded proteins. (2) Expression of proteins involved in polypeptide folding is strongly enhanced by a process called the Unfolded Protein Response (UPR). (3) Proteins missing the proper tertiary structure are degraded by the ER-Associated protein Degradation (ERAD) mechanism.

 

Recent studies in S. cerevisiae have shown that the processes of UPR and ERAD are functionally linked to each other. Cells lacking a functional ERAD show a constitutive activation of UPR. In addition, many of the components of ERAD are under the direct transcriptional control of UPR. Finally, while neither UPR nor ERAD are essential for cell viability, deletion of both pathways results in severe growth impairment.

 

UPR and ERAD are conserved between yeast and mammalian cells. One of the components of mammalian UPR is the protease presenilin-1. Mutations in the gene for presenilin-1 cause early-onset familial Alzheimer’s disease. Interestingly, inhibition of proteolysis by the ubiquitin-26S proteasome system has also been described for Alzheimer’s disease. This suggests a link between UPR and ERAD in mammalian cells.

 

The recently identified gene Mif1 is a possible candidate to form a direct link between UPR and ERAD in mammalian cells. The Mif1 gene is under the direct control of UPR. Mif1 is a trans-ER-membrane protein, with both the N- and the C-termini facing the cytoplasmic side of the ER membrane. It contains an N-terminal ubiquitin-like domain. It is anticipated that Mif1 may associate through its ubiquitin-like domain with the 26S proteasome, in this way connecting the protein degradation machinery to the ER membrane and resulting in an efficient ERAD.