Applicability of Instability Index for In vitro Protein Stability Prediction

Protein and Peptide Letters
Dilani G GamageTimothy G Russell

Abstract

The dipeptide composition-based Instability Index (II) is one of the protein primary structure-dependent methods available for in vivo protein stability predictions. As per this method, proteins with II value below 40 are stable proteins. Intracellular protein stability principles guided the original development of the II method. However, the use of the II method for in vitro protein stability predictions raises questions about the validity of applying the II method under experimental conditions that are different from the in vivo setting. The aim of this study is to experimentally test the validity of the use of II as an in vitro protein stability predictor. A representative protein CCM (CCM - Caulobacter crescentus metalloprotein) that rapidly degrades under in vitro conditions was used to probe the dipeptide sequence-dependent degradation properties of CCM by generating CCM mutants to represent stable and unstable II values. A comparative degradation analysis was carried out under in vitro conditions using wildtype CCM, CCM mutants and two other candidate proteins: metallo-β-lactamase L1 and α -S1- casein representing stable, borderline stable/unstable, and unstable proteins as per the II predictions. The effect of temperatu...Continue Reading

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Citations

Jun 10, 2020·International Journal of Molecular Sciences·Mohamad Farhan Mohamad SobriNorhayati Ramli
Jul 12, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Olga D NovikovaOlga L Voronina
Nov 15, 2020·Expert Opinion on Therapeutic Patents·Giannamaria Annunziato, Gabriele Costantino
Mar 9, 2021·Applied Biochemistry and Biotechnology·Debanjan Mitra, Pradeep K Das Mohapatra
Jul 25, 2021·International Journal of Biological Macromolecules·Dielson da S VieiraFrancisco L F Feitosa
Oct 1, 2021·Journal, Genetic Engineering & Biotechnology·Rahmat Eko SanjayaNi Nyoman Tri Puspaningsih

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