Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from Solanum tuberosum subsp. andigenum cv. Churqueña

International Journal of Molecular Sciences
Juliana CotabarrenWalter David Obregón

Abstract

Cystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three f...Continue Reading

References

Apr 1, 1989·Archives of Biochemistry and Biophysics·G A HomandbergR J Peanasky
Sep 25, 1982·Journal of Molecular Biology·D C Rees, W N Lipscomb
Dec 31, 1980·Biochemical and Biophysical Research Communications·G M Hass, C A Ryan
Apr 14, 1981·Biochemistry·G M Hass, M A Hermodson
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Oct 29, 1998·Toxicon : Official Journal of the International Society on Toxinology·R S Norton, P K Pallaghy
Nov 25, 1998·Journal of Molecular Biology·J M MasF X Avilés
Aug 4, 1999·Proceedings of the National Academy of Sciences of the United States of America·J P TamK W Chiu
Jan 20, 2006·Journal of Thrombosis and Haemostasis : JTH·X WangW A Schumacher
Feb 3, 2006·Protein Science : a Publication of the Protein Society·Cyril PimentelHervé Darbon
Feb 22, 2007·Current Pharmaceutical Design·Joan L ArolasLloyd D Fricker
Apr 5, 2007·Nature Protocols·Hermann Schägger
Jun 6, 2007·Plant Biotechnology Journal·Jordi QuilisBlanca San Segundo
Jul 27, 2007·Journal of Insect Physiology·Haiyan GongKozo Fujisaki
Nov 21, 2007·Nucleic Acids Research·Jérôme GracyLaurent Chiche
Jun 4, 2008·Expert Opinion on Pharmacotherapy·Jason A WilliamsJames E Heavner
Jan 31, 2009·Proceedings of the National Academy of Sciences of the United States of America·Laura SanglasJoan L Arolas
Dec 17, 2009·Protein and Peptide Letters·W D ObregónN S Priolo
Feb 24, 2010·Journal of Pharmacological and Toxicological Methods·R RinkG N Moll
Dec 24, 2010·British Journal of Pharmacology·C CavalliniT Pandolfini
Sep 28, 2014·Molecular Genetics and Genomics : MGG·Matthias FischerChristiane Gebhardt
Aug 19, 2015·Biotechnology Advances·Atanas G AtanasovHermann Stuppner
Nov 4, 2015·Nucleic Acids Research·Neil D RawlingsRobert Finn

❮ Previous
Next ❯

Citations

Feb 2, 2020·Plant Science : an International Journal of Experimental Plant Biology·Juliana CotabarrenWalter David Obregón

❮ Previous
Next ❯

Datasets Mentioned

BETA
MF682092

Methods Mentioned

BETA
thermal treatment
electrophoresis
PCR
size-exclusion chromatography
Affinity Chromatography

Software Mentioned

MASCOT
BioTools
BLAST

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.