Conformation and Stability of Recombinant HIV-1 Capsid Protein P24 (rp24)

Misselwitz, R.; Hausdorf, G.; Welfle, K.; Höhne, W. E.; Welfle, H.

doi:10.1007/978-94-011-0371-8_46

Cited by 8 publications

(11 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…). The distribution of the secondary structural elements is in good agreement with results published previously for the His 6 ‐tagged CA at pH 7.5 , and also with the predictions made from the amino acid sequence of the wild‐type protein (Table ). The spectra of A78V and L189F mutants showed similar features to that of the wild‐type, indicating that these proteins have the same distribution of secondary structural elements (Table ).…”

Section: Resultssupporting

confidence: 90%

“…CD spectra were analyzed by the CDSSTR analysis program; results are summarized in Table . These spectra were measured at pH 7.5 in which the CA protein maintains its structural integrity .…”

Section: Resultsmentioning

confidence: 99%

“…NMR spectroscopic and X‐ray crystallographic studies revealed that the domain organization of the HIV‐1 CA protein is highly α‐helical. Moreover, circular dichroism spectroscopic (CD) study of a recombinant p24 protein (r‐p24) showed that under native conditions, 56% of the protein is folded into α‐helices, 3% into β‐sheets, and 20% into turns .…”

mentioning

confidence: 99%

See 2 more Smart Citations

Effect of internal cleavage site mutations in human immunodeficiency virus type 1 capsid protein on its structure and function

et al. 2016

View full text Add to dashboard Cite

The capsid protein of the human immunodeficiency virus type 1 has been found to be a substrate of the retroviral protease in vitro , and its processing was predicted to be strongly dependent on a pH ‐induced conformational change. Several protease cleavage sites have been identified within the capsid protein, but the importance of its cleavage by the viral protease at the early phase of infection is controversial. To confirm the relevance of this process, we aimed to design, produce, and characterize mutant capsid proteins, in which the protein susceptibility toward HIV ‐1 protease is altered without affecting other steps of the viral life cycle. Our results indicate that while the introduced mutations changed the cleavage rate at the mutated sites of the capsid protein by HIV ‐1 protease, some of them caused only negligible or moderate structural changes (A78V, L189F, and L189I). However, the effects of other mutations (W23A, A77P, and L189P) were dramatic, as assessed by secondary structure determination or cyclophilin A‐binding assay. Based on our observations, the L189F mutant capsid remains structurally and functionally unchanged and may therefore be the best candidate for use in studies aimed at better understanding the role of the protease in the early postentry events of viral infection or retrovirus‐mediated gene transduction.

show abstract

Section: Resultssupporting

confidence: 90%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Effect of internal cleavage site mutations in human immunodeficiency virus type 1 capsid protein on its structure and function

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Circular Dichroism. CD measurements were performed with a Jasco J-720 spectropolarimeter (Jasco Instruments S. A., Japan) as described recently (Misselwitz et al, 1995). The instrument was equipped with a thermostated cell holder and a temperature control system (Neslab, USA).…”

Section: Methodsmentioning

confidence: 99%

C Repeats of the Streptococcal M1 Protein Achieve the Human Serum Albumin Binding Ability by Flanking Regions Which Stabilize the Coiled-Coil Conformation

Gubbe¹,

Misselwitz

Welfle

et al. 1997

Biochemistry

View full text Add to dashboard Cite

The M and M-like proteins of Streptococcus pyogenes are fibrous cell surface proteins. They have multiple binding sites for several human proteins and are composed of the C-terminal anchor domain, the alpha-helical coiled-coil domain, and the N-terminal non-coiled-coil domain. The coiled-coil domain of the M1 protein consists of repeat units called B, C, and D and a spacer unit S between B and C. Recombinant fragments A-B-S-C-D, A-B-S, B-S-C, S-C, S-C-D, C-D, and C of the coiled-coil domain were studied by analyzing their secondary structures and binding affinities to human serum albumin (HSA). As shown by circular dichroism, all fragments are in an alpha-helical conformation. C-D and S-C-D form coiled coils at room temperature and bind below 37 degrees C with high affinity to HSA. C-D and S-C-D unfold in two steps with Tm values of approximately 31 and approximately 65 degrees C; complex formation with HSA increases the unfolding temperatures. B-S-C has a lower alpha-helical content, a less pronounced coiled-coil conformation, and a reduced thermal stability, binds HSA weaker, and is only slightly stabilized by HSA binding in comparison to C-D and S-C-D. C and S-C are less stable than the other fragments and are not organized as coiled coils showing some features of alpha-helical single strands only below 20 degrees C, and binding of HSA was not observed. The results indicate that the formation of coiled-coil structures, supported by flanking D regions and, to a lesser extent also B regions, is essential for the binding of C repeat units to HSA.

show abstract

“…Considering that p24 protein has a predominant ahelical structure as was previously determined by CD studies [32], we focused our attention on the variations of a-helix content in the different peptides by means of CD analyses. The presence in p24-1n of several amino acid residues close to the N-terminal end with a high propensity to adopt a helical structure (Glu, Met, Ala, Leu,Lys) that were absent from the modified sequences (p24-1ml and p24-1mc), agrees with the CD results.…”

Section: Discussionmentioning

confidence: 99%

Secondary structure-improved bioaffinity correlation in elongated and modified synthetic epitope peptides from p24 HIV-1 core protein

Tonarelli¹,

Lottersberger²,

Salvetti³

et al. 2000

Lett Pept Sci

View full text Add to dashboard Cite

SummaryIn order to study the correlation between secondary structure and bioafflnity of long and modified sequences of p24 protein from HIV-1, three peptides containing the minimal size epitope from region 208-217 (EAAEWDRVHP) were prepared. It was found that peptide p24-1n, an elongated native sequence, has the lowest KD and a predominant a-helix structure in the presence of trifluoroethanol. Three peptides containing another epitope from region 293-302 (FRDYVDRFK) were also synthesized. We have observed that modifications on the native sequence p24-2n (region 287-308) increased the a-helix content and this was correlated with an improvement of the recognition by antibodies in ELISA.Abbreviations: CD, circular dichroism; DMF, N,N-dimethylformamide; DMSO, dimethyl sulfoxide; ELISA, enzyme linked immunosorbent assay; EDT, 1,2-ethanedithiol; Fmoc, fluorenylmethoxycarbonyl; FAB-MS, fast atom bombardment mass spectrometry; HPLC, high pressure liquid chromatography; MeCN, acetonitrile; NMM, N-methylmorpholine; PyBOP, benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate; SELCOM, self consistent method; TMB, 3,3t,5,51-tetramethylbenzidine; TFA, trifluoroacetic acid; TFE, trifluoroethanol; Aminoacid symbols denote the L-configurations. Abbreviations for amino acids and nomenclature of peptides follow the recommendations of the IUPAC-IUB Commission on Biochemical Nomenclature (Eur. J. Biochem., 138 (1984) 9).

show abstract

Conformation and Stability of Recombinant HIV-1 Capsid Protein P24 (rp24)

Cited by 8 publications

References 0 publications

Effect of internal cleavage site mutations in human immunodeficiency virus type 1 capsid protein on its structure and function

Effect of internal cleavage site mutations in human immunodeficiency virus type 1 capsid protein on its structure and function

C Repeats of the Streptococcal M1 Protein Achieve the Human Serum Albumin Binding Ability by Flanking Regions Which Stabilize the Coiled-Coil Conformation

Secondary structure-improved bioaffinity correlation in elongated and modified synthetic epitope peptides from p24 HIV-1 core protein

Contact Info

Product

Resources

About