Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2012 Jul 18;7(7):e40486. doi: 10.1371/journal.pone.0040486

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Clark et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

PMC Copyright notice

Samples of Hemoglobin A or Hemoglobin S were heated in the presence and absence of αB crystallin and then the interactive aggregates or bound complexes were separated by centrifugation. At low speed (2500 g) only the largest aggregates were in the insoluble (I) fraction, and the Hb bound to αB crystallin remained soluble in the supernatant (soluble) fraction. The soluble fraction was centrifuged again at 14,000 g using a 100 kDa spin-filter to separate the HbA or HbS bound to αB crystallin in the soluble concentrate (Sc) from the soluble HbA or HbS in the filtrate (Sf). With thermal destabilization, aggregation was expected to increase the amount of αB crystallin bound HbA or HbS in the insoluble (I) and soluble concentrate (Sc) fractions and decrease the amount in the soluble filtrate (Sf) fraction. The centrifugation experiments were conducted under conditions of minimal thermal stress to determine the sensitivity of αB crystallin to the earliest stages of protein unfolding and aggregation.