Minimization of a eukaryotic mini-intein

Elleuche, Skander; Döring, Kristin; Pöggeler, Stefanie

doi:10.1016/j.bbrc.2007.11.126

Cited by 20 publications

(13 citation statements)

References 32 publications

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“…(37). The PRP8 mini-intein encoding plasmid was kindly provided by Stephanie Pöggeler (Georg August Universität, Göttingen, Germany) (38). The GFP encoding sequence was amplified from pEGFP-C1 (Clontech), which encodes a modified Aequorea victoria GFP.…”

Section: Methodsmentioning

confidence: 99%

Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

Farfán-Arribas

Stern

Rock

2012

Proc. Natl. Acad. Sci. U.S.A.

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All vertebrate nucleated cells generate peptides from their expressed gene products and then display them at the cell surface bound to MHC class I molecules. This allows CD8 + T cells to detect and eliminate abnormal cells that are synthesizing foreign proteins, e.g., from viruses or mutations. To permit the immune system to more uniformly monitor a cell's proteins, regardless of their half-life or location, it has been thought that the products of rapid degradation of the mistakes of protein synthesis (defective ribosomal products, DRiPs) preferentially contribute to the class I-presented peptides. However, using intein catalysis to generate peptide sequences exclusively by posttranslational splicing of mature proteins, we show here that presented peptides can be generated from fully folded and functional proteins. Remarkably, the presentation of peptides from two model mature proteins is just as efficient as from newly synthesized proteins subject to errors in translation or folding. These results indicate that for the constructs we have analyzed, DRiPs are not a more efficient source of class I peptides for antigen presentation than the turnover of mature functional proteins. Accordingly, our data suggest that one of the major ways the immune system evaluates the health of cells is by monitoring the breakdown products of the proteome.

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Section: Methodsmentioning

confidence: 99%

Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

Farfán-Arribas

Stern

Rock

2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Site-directed mutations R104L, E107D, and S130T were introduced using an inverse PCR procedure with Phusion TM High-Fidelity DNA Polymerase (New England Biolabs, Inc.) as described previously (Elleuche et al, 2008). In these cases all forward primers contained the exchanged nucleotides to substitute the defined amino-acids (Table S1).…”

Section: Site-directed Mutagenesis Of the His-tagged S Macrospora Cyn1mentioning

confidence: 99%

A cyanase is transcriptionally regulated by arginine and involved in cyanate decomposition in Sordaria macrospora

Elleuche

Pöggeler

2008

Fungal Genetics and Biology

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“…Other inteins are naturally occurring mini-inteins that are as small as 134 residues and lack an endonuclease domain (20). Studies of both native and engineered mini-inteins helped define the intein splicing domain (20, 23–27). Intein genes may also be split between motifs B and F; however, the expressed precursor protein fragments rapidly assemble to splice in trans by the same mechanisms used in cis -splicing inteins (28–32).…”

Section: Introductionmentioning

confidence: 99%

Protein Splicing: How Inteins Escape from Precursor Proteins

Mills

Johnson

Perler

2014

Journal of Biological Chemistry

121

140

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Inteins are nature's escape artists; they facilitate their excision from flanking polypeptides (exteins) concomitant with extein ligation to produce a mature host protein. Splicing requires sequential nucleophilic displacement reactions catalyzed by strategies similar to proteases and asparagine lyases. Inteins require precise reaction coordination rather than rapid turnover or tight substrate binding because they are single turnover enzymes with covalently linked substrates. This has allowed inteins to explore alternative mechanisms with different steps or to use different methods for activation and coordination of the steps. Pressing issues include understanding the underlying details of catalysis and how the splicing steps are controlled.

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Minimization of a eukaryotic mini-intein

Cited by 20 publications

References 32 publications

Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

A cyanase is transcriptionally regulated by arginine and involved in cyanate decomposition in Sordaria macrospora

Protein Splicing: How Inteins Escape from Precursor Proteins

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