Abstract:
PURPOSE:The prevalence of cataract increases with age. Conversely, the abundance of native α-crystallin diminishes with age and cataract development. We hypothesize replenishing lens α-crystallin may delay or prevent cataract. Herein we investigated the ability of cell penetration peptides (CPP) to enhance entry of α-crystallins into lens-derived cells. METHODS:Recombinant αB-crystallins were modified by the addition of CPPs. Candidate CPP were designed with reference to the HSV-1 glycoprotein C gene (gC) or the HIV-1 TAT peptide. αB-crystallins produced by fusing gC or TAT were over-expressed in E. coli. Purified proteins were subjected to size exclusion chromatography (SEC) to characterize oligomeric complexes (OC). Chaperone-like activity (CLA) was evaluated by measuring the ability of α-crystallins to suppress chemically-induced protein aggregation. To evaluate protein uptake, labeled α-crystallins were incubated with HLE B3 cells and monitored by fluorescence microscopy for 48 hours. RESULTS:We examined the effects of the addition of CPP on the structure, CLA, and cell transduction properties of αB-crystallins. C-terminal CPP fused crystallins had poor solubility. In contrast, N-terminal tagged αB-crystallins were soluble. These modified αB-crystallins formed OC that were larger than wild-type based on SEC. Wild-type and gC tagged αB-crystallin displayed robust CLA. Subunit exchange was observed when gC-fused αB-crystallin was mixed with αA. In contrast to wild-type, modified α-crystallins accumulated in HLE B3 cells. CONCLUSIONS:Addition of CPP improves the uptake of αB-crystallins into HLE B3 cells. No undesirable changes to the chaperone-like abilities of α-crystallins were observed in αB-crystallin modified by the addition of the gC-derived CPP.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Mueller NH,Ammar DA,Petrash JMdoi
10.1167/iovs.12-10947subject
Has Abstractpub_date
2013-01-02 00:00:00pages
2-8issue
1eissn
0146-0404issn
1552-5783pii
iovs.12-10947journal_volume
54pub_type
杂志文章abstract:PURPOSE:To demonstrate the potential use of subpixel image processing methods to perform automated Hirschberg measurements of strabismic deviations using relatively inexpensive personal computer hardware; to determine if the method might allow screening for strabismus using full-face video images obtained from a distan...
journal_title:Investigative ophthalmology & visual science
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abstract:Purpose:To compare blood pressure and urine albumin-creatinine ratio over time for participants receiving aflibercept, bevacizumab, or ranibizumab. Methods:Preplanned secondary analyses from a randomized trial comparing aflibercept, bevacizumab, and ranibizumab for diabetic macular edema (DME). The Diabetic Retinopath...
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journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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更新日期:1999-08-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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doi:
更新日期:2001-04-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:2000-11-01 00:00:00
abstract::An imaging fundus reflectometer has been used to study the distribution and regeneration of visual pigments in the retina of the adult living cat. Measurements were made over an area of tapetal retina extending 25 degrees nasal and 40 degrees temporal to the area centralis on the horizontal meridian and 5 degrees infe...
journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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journal_title:Investigative ophthalmology & visual science
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更新日期:1982-02-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
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更新日期:1978-12-01 00:00:00
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