Identification of C-terminal extensions that protect proteins from intracellular proteolysis.

Revertants of defective mutants in the Arc repressor of bacteriophage P22 were isolated. Five of the six reverting mutations were frameshifts near the end of the coding sequence which resulted in proteins with C-terminal extensions. Each of the reverting mutations prolong the half-lives in vivo of the proteins in which they reside, yet they do not alter the thermodynamic stability, structure, oligomeric form, or DNA-binding properties of these proteins. Fusion of one of these tails to the C-terminal end of a mutant form of the N-terminal domain of lambda repressor also prevented proteolysis of this protein. These C-terminal sequences may prevent degradation by blocking the recognition of unstable proteins by the proteolytic machinery in the cell.

• Publication year

  1989

• Venue

  Journal of Biological Chemistry

• Publication date

  1989-05-05

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/s0021-9258(18)83276-8PMID 2651443
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• The structural stability of a protein is an important determinant of its proteolytic susceptibility in Escherichia coli.

  1989cited by this paper
• Interaction of the bacteriophage P22 Arc repressor with operator DNA.

  1987cited by this paper
• Comparison of the subsite specificity of the mammalian neutral endopeptidase 24.11 (enkephalinase) to the bacterial neutral endopeptidase thermolysin.

  1986cited by this paper
• The bacteriophage P22 arc and mnt repressors. Overproduction, purification, and properties.

  1985cited by this paper
• Lambda repressor mutations that increase the affinity and specificity of operator binding.

  1985cited by this paper
• A gene regulating the heat shock response in Escherichia coli also affects proteolysis.

  1984cited by this paper
• Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli

  1984cited by this paper
• Cloning vectors that yield high levels of single-stranded DNA for rapid DNA sequencing.

  1984cited by this paper
• Studies on transformation of Escherichia coli with plasmids.

  1983cited by this paper
• On the determination of deoxyribonucleic acid-protein interaction parameters using the nitrocellulose filter-binding assay.

  1983cited by this paper
• Control of phage lambda development by stability and synthesis of cII protein: role of the viral cIII and host hflA, himA and himD genes.

  1982cited by this paper
• Sequence determinants of promoter activity.

  1982cited by this paper
• A plasmid cloning vector for the direct selection of strains carrying recombinant plasmids.

  1981cited by this paper
• Advanced bacterial genetics

  1980cited by this paper
• Deg phenotype of Escherichia coli lon mutants

  1978cited by this paper
• DNA sequencing with chain-terminating inhibitors.

  1977cited by this paper
• Experiments in molecular genetics

  1972cited by this paper

• Contribution of a C-Terminal Extension to the Substrate Affinity and Oligomeric Stability of Aldehyde Dehydrogenase from Thermus thermophilus HB27

  2024cites this paper
• Viewing Protein Fitness Landscapes Through a Next-Gen Lens

  2014cites this paper
• High-quality combinatorial protein libraries using the binary patterning approach.

  2014cites this paper
• Deletion of the carboxyl‐terminal residue disrupts the amino‐terminal folding, self‐association, and thermal stability of an amphipathic antimicrobial peptide

  2014cites this paper
• Overview of Approaches to Preventing and Avoiding Proteolysis During Expression and Purification of Proteins

  2013cites this paper
• Mutagenic dissection of the sequence determinants of protein folding, recognition, and machine function

  2013cites this paper
• Design and Screening of a Glial Cell-Specific, Cell Penetrating Peptide for Therapeutic Applications in Multiple Sclerosis

  2012cites this paper
• Protein Folding and Solubility: Pathways and High‐Throughput Assays

  2011cites this paper
• Alleviation of Proteolytic Sensitivity To Enhance Recombinant Lipase Production in Escherichia coli

  2009cites this paper
• Conformation of a plasmid replication initiator protein affects its proteolysis by ClpXP system

  2009cites this paper
• Molecular and Genetic Strategies to Enhance Functional Expression of Recombinant Protein in Escherichia coli

  2009cites this paper
• Intracellular proteolysis: Signals of selective protein degradation

  2007cites this paper
• Stress-driven in vivo selection of a functional mini-gene from a randomized DNA library expressing combinatorial peptides in Escherichia coli.

  2007cites this paper
• Alanine racemase from the acidophile Acetobacter aceti.

  2007cites this paper
• De novo proteins from binary-patterned combinatorial libraries.

  2006cites this paper
• Protein Design

  2006cites this paper
• Protein Engineering Protocols

  2006cites this paper
• Substrate denaturation and translocation by a proteolytic machine

  2005cites this paper
• Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli

  2005cites this paper
• Proteases and protein degradation inEscherichia coli

  2005cites this paper
• Combinatorial approaches to protein stability and structure.

  2004cites this paper
• Brucella abortus RB51 vaccine: Testing its Spectrum of Protective and Curative Characteristics

  2004cites this paper
• BgK, a disulfide-containing sea anemone toxin blocking K+ channels, can be produced in Escherichia coli cytoplasm as a functional tagged protein.

  2004cites this paper
• DIMACS Workshop on Evolution as Computation Toward in vivo Digital Circuits

  2004cites this paper
• Sculpting the proteome with AAA(+) proteases and disassembly machines.

  2004cites this paper
• Deletion of the prc (tsp) gene provides evidence for additional tail-specific proteolytic activity in Escherichia coli K-12

  2004cites this paper
• Regulation by Proteolysis: Energy-Dependent Proteases

  2003cites this paper
• Toward in vivo Digital Circuits

  2002cites this paper
• Extra terminal residues have a profound effect on the folding and solubility of a Plasmodium falciparum sexual stage-specific protein over-expressed in Escherichia coli.

  2002cites this paper
• P22 Arc repressor: Enhanced expression mutants by addition of polar C-terminal of unstable sequences

  2002cites this paper
• Role of an N(cap) residue in determining the stability and operator-binding affinity of Arc repressor.

  2002cites this paper
• Cellular computation and communications using engineered genetic regulatory networks

  2001cites this paper
• Stabilizing C‐terminal tails on AraC

  2001cites this paper
• [Energy-dependent selective intracellular proteolysis. Structure, active sites and specificity of ATP-dependent proteinases].

  2001cites this paper
• Transcriptional repressor CopR: dissection of stabilizing motifs within the C terminus.

  2001cites this paper
• Striking stabilization of Arc repressor by an engineered disulfide bond.

  2000cites this paper
• Modification of the carboxy-terminal amino acid sequence alters the Escherichia coli expression of a gene encoding multiple repeats of a bovine growth hormone releasing factor analog.

  2000cites this paper
• Core-Binding Factor Influences the Disease Specificity of Moloney Murine Leukemia Virus

  1999cites this paper
• P1 ParB Domain Structure Includes Two Independent Multimerization Domains

  1999cites this paper
• A Mutant HemA Protein with Positive Charge Close to the N Terminus Is Stabilized against Heme-Regulated Proteolysis in Salmonella typhimurium

  1999cites this paper
• Origins of DNA-binding specificity: role of protein contacts with the DNA backbone.

  1999cites this paper
• Tolerance of Arc repressor to multiple-alanine substitutions.

  1999cites this paper
• Lon-mediated proteolysis of the Escherichia coli UmuD mutagenesis protein: in vitro degradation and identification of residues required for proteolysis.

  1998cites this paper
• The ClpXP and ClpAP proteases degrade proteins with carboxy-terminal peptide tails added by the SsrA-tagging system.

  1998cites this paper
• Regulation of protein degradation in plants.

  1997cites this paper
• Covalent attachment of Arc repressor subunits by a peptide linker enhances affinity for operator DNA.

  1996cites this paper
• The CBFbeta subunit is essential for CBFalpha2 (AML1) function in vivo.

  1996cites this paper
• Strategies for achieving high-level expression of genes in Escherichia coli.

  1996cites this paper
• Sequence Determinants of C-terminal Substrate Recognition by the Tsp Protease (*)

  1996cites this paper
• Upstream strategies to minimize proteolytic degradation upon recombinant production in Escherichia coli.

  1996cites this paper
• The position of the heterologous domain can influence the solubility and proteolysis of beta-galactosidase fusion proteins in E. coli.

  1996cites this paper
• Proteases and their targets in Escherichia coli.

  1996cites this paper
• The CBFb subunit is essential for CBFa2 (AML1)function in vivo

  1996cites this paper
• P22 Arc repressor: role of cooperativity in repression and binding to operators with altered half-site spacing.

  1995cites this paper
• Critical side-chain interactions at a subunit interface in the Arc repressor dimer.

  1995cites this paper
• Dramatic changes in DNA-binding specificity caused by single residue substitutions in an Arc/Mnt hybrid repressor

  1995cites this paper
• C‐terminal specific protein degradation: Activity and substrate specificity of the Tsp protease

  1995cites this paper
• Crystal structure, folding, and operator binding of the hyperstable Arc repressor mutant PL8.

  1995cites this paper
• Are buried salt bridges important for protein stability and conformational specificity?

  1995cites this paper
• P22 Arc repressor: folding kinetics of a single-domain, dimeric protein.

  1994cites this paper
• Protein design by binary patterning of polar and nonpolar amino acids.

  1993cites this paper
• P22 Arc repressor: Enhanced expression of unstable mutants by addition of polar C‐terminal sequences

  1993cites this paper
• Baculovirus-mediated expression of rat liver CTP:phosphocholine cytidylyltransferase.

  1993cites this paper
• Regulation by proteolysis: energy-dependent proteases and their targets

  1992cites this paper
• Structural characteristics of an abnormal protein influencing its proteolytic susceptibility.

  1992cites this paper
• Shuttle vectors conferring hygromycin B resistance to E. coli and to mammalian cells. Differential expression of carboxyterminal fusion proteins.

  1992cites this paper
• Tsp: a tail-specific protease that selectively degrades proteins with nonpolar C termini.

  1992cites this paper
• Proteases and protein degradation in Escherichia coli.

  1992cites this paper
• Stabilization of recombinant proteins from proteolytic degradation in Escherichia coli using a dual affinity fusion strategy

  1991cites this paper
• In vivo interaction of Escherichia coli lac repressor N-terminal fragments with the lac operator.

  1991cites this paper
• Identification of three amino acid residues in the B subunit of Shiga toxin and Shiga-like toxin type II that are essential for holotoxin activity

  1991cites this paper
• Mutation in the Escherichia coli htpR locus results in stabilization of recombinant expression products that are susceptible to proteolytic degradation.

  1991cites this paper
• A streptomycin selection for DNA-binding activity.

  1991cites this paper
• Proper and improper folding of proteins in the cellular environment.

  1991cites this paper
• Random mutagenesis of protein sequences using oligonucleotide cassettes.

  1991cites this paper
• Arc repressor is tetrameric when bound to operator DNA.

  1990cites this paper
• Differential stability of two apo-isocytochromes c in the yeast Saccharomyces cerevisiae.

  1990cites this paper
• Role of the Cro repressor carboxy-terminal domain and flexible dimer linkage in operator and nonspecific DNA binding.

  1990cites this paper
• Carboxy-terminal determinants of intracellular protein degradation.

  1990cites this paper
• Identification of key residues in the amino-terminal third of human interleukin-1 alpha.

  1990cites this paper
• An essential proline in lambda repressor is required for resistance to intracellular proteolysis.

  1990cites this paper
• Minimizing proteolysis in Escherichia coli: genetic solutions.

  1990cites this paper
• Reverse hydrophobic effects relieved by amino-acid substitutions at a protein surface

  1990cites this paper
• The structural stability of a protein is an important determinant of its proteolytic susceptibility in Escherichia coli.

  1989cites this paper
• Genetics of proteolysis in Escherichia coli*.

  1989cites this paper
• Genetic analysis of protein stability and function.

  1989cites this paper
• Equilibrium dissociation and unfolding of the Arc repressor dimer.

  1989cites this paper
• Induction of a heat shock-like response by unfolded protein in Escherichia coli: dependence on protein level not protein degradation.

  1989cites this paper
• Amino acid substitutions that increase the thermal stability of the lambda Cro protein.

  1989cites this paper
• Amino acid substitutions that increase the thermal stability of the λ Cro protein

  1989cites this paper