Biochemical characterization of a novel α/β-hydrolase/FSH from the white shrimp Litopenaeus vannamei

Karina D. Garcia-Orozco, Francisco Cinco-Moroyoqui, Lucía T. Angulo-Sanchez, Enrique Marquez-Rios, Armando Burgos-Hernandez, Jose L. Cardenas-Lopez, Carolina Gomez-Aguilar, David O. Corona-Martinez, Gloria Saab-Rincon, Rogerio R. Sotelo-Mundo

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Energy buffering systems are key for homeostasis during variations in energy supply. Spiders are the most important predators for insects and therefore key in terrestrial ecosystems. From biomedical interest, spiders are important for their venoms and as a source of potent allergens, such as arginine kinase (AK, EC 2.7.3.3). AK is an enzyme crucial for energy metabolism, keeping the pool of phosphagens in invertebrates, and also an allergen for humans. In this work, we studied AK from the Argentininan spider Polybetes pythagoricus (PpAK), from its complementary DNA to the crystal structure. The PpAK cDNA from muscle was cloned, and it is comprised of 1068 nucleotides that encode a 384-amino acids protein, similar to other invertebrate AKs. The apparent Michaelis-Menten kinetic constant (Km) was 1.7 mM with a kcat of 75 s−1. Two crystal structures are presented, the apoPvAK and PpAK bound to arginine, both in the open conformation with the active site lid (residues 310–320) completely disordered. The guanidino group binding site in the apo structure appears to be organized to accept the arginine substrate. Finally, these results contribute to knowledge of mechanistic details of the function of arginine kinase.

Original languageEnglish
Article number674
JournalBiomolecules
Volume9
Issue number11
DOIs
StatePublished - 1 Nov 2019

Bibliographical note

Funding Information:
Funding: This research was funded by CONACYT grant CB-2014-237963 and INFR-2014-01-225455. Additionally, by grant UNAM-CIC-CIAD-2018 and -2019 for academic exchange.

Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Allergen
  • Arachnida
  • Arginine kinase
  • Arthropoda
  • CDNA cloning
  • Crystal structure
  • Open conformation
  • Phosphagen
  • Polybetes pytagoricus
  • Spider

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