Two contiguous thrombin fragments of human somatotropin form a functionally active recombinant, but the two homologous fragments from sheep hormone do not

Gráf, László; Li, Choh Hao; Chen, Cheng; Jibson, Michael D.

doi:10.1021/bi00528a031

Cited by 18 publications

(14 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The two-chain form, however, resembles hGH far more closely when examined by c.d. (Graf et al, 1981). These observations are in agreement with earlier reports suggesting that most of the antigenicity of the hGH molecule is retained in the first 134 residues from the amino terminus (Clarke et al, 1974;Niall et al, 1971).…”

Section: Discussionsupporting

confidence: 93%

“…It has also been demonstrated that all tested monoclonal antibodies with combining site specificities towards four distinct epitopes which are expressed on intact hGH molecules (Ivanyi, 1982a(Ivanyi, , 1982b (Graf et al, 1981). The two-chain form, however, resembles hGH far more closely when examined by c.d.…”

Section: Discussionmentioning

confidence: 98%

“…Digestion of hGH with plasmin, thrombin or subtilisin (Li and Bewley, 1976; Lewis et al, 1980;Graf et al, 1981) leads to cleavage in the sequence region 134-149, resulting in a two-chain form which is conformationally similar to the un-*To whom reprint requests should be sent.…”

Section: Introductionmentioning

confidence: 99%

“…digested molecule (Graf et al, 1981). The two-chain form generated by plasmin cleavage consists of residues 1 -134 and 141 -191 linked by a single disulphide bond between residues 53 and 165, whereas subtilisin digestion results in a mixture of three major cleaved forms which arise from a 'nick' at residues 139, 146 and 149 (hGH-S) (Lewis et al, 1977).…”

Section: Introductionmentioning

confidence: 99%

“…The two-chain form generated by plasmin cleavage consists of residues 1 -134 and 141 -191 linked by a single disulphide bond between residues 53 and 165, whereas subtilisin digestion results in a mixture of three major cleaved forms which arise from a 'nick' at residues 139, 146 and 149 (hGH-S) (Lewis et al, 1977). Digestion of hGH with thrombin results in a more specific hydrolysis giving a single cleavage and producing a hormone representing the full primary structure of the active hormone (residues 1 -134 and 135 -191) (Graf et al, 1981). The N-terminal, 15-K fragment of hGH (residues 1-134), derived from either plasmin or subtilisin digestion, retains activity in the pigeon crop-sac and rat tibia tests, whereas the C-terminal 7-K fragment is almost devoid of biological activity Lewis et al, 1977).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Antigenic, receptor-binding and mitogenic activity of proteolytic fragments of human growth hormone.

Aston¹,

Iványi²

1983

The EMBO Journal

View full text Add to dashboard Cite

Analysis of the subtilisin-digested, two-chain form of human growth hormone (hGH) and its constituent polypeptide fragments has been aided by the use of monoclonal antibodies which bind specifically to four distinct epitopes on the native hormone. Using the SDS-polyacrylamide immunoblotting technique, it was shown that one epitope (shared with human chorionic somatomammotropin) detected by EB1 (or EB3) antibody was expressed to a similar extent by both the N-terminal (15 K) and C-terminal (7 K) polypeptides. This epitope is unique in that it represents a repeating determinant within the single chain structure of the hormone. Another three epitopes detected by monoclonal antibodies QA68/NA27, NA71 or NA39/EB2 were absent from the 7-K fragment but were expressed on the 15-K fragment to a similar extent to that on unmodified growth hormone. Binding of NA71 antibody was demonstrated only by radioimmunoassay since this, presumably conformational epitope could not be detected by immunoblotting. The functional importance of the 15-K peptide was demonstrated by its ability to bind specifically to hormone receptors on IM9 human lymphoblastoid cells and by its retention of mitogenicity for the NB2 rat lymphoma ceil line. However, all tested monoclonal antibodies inhibited the binding of [1251]15-K to IM9 cell receptors by either steric hindrance or by an allosteric mechanism and therefore could not be further related topographically to the receptor-binding moiety of hGH.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Antigenic, receptor-binding and mitogenic activity of proteolytic fragments of human growth hormone.

Aston¹,

Iványi²

1983

The EMBO Journal

View full text Add to dashboard Cite

show abstract

Prediction of protein helix content from an autocorrelation analysis of sequence hydrophobicities

Horne

1988

Biopolymers

View full text Add to dashboard Cite

SynopsisI t is demonstrated that protein a-helix content can be predicted from an autocorrelation analysis of the protein hydrophobicity sequence. The Fourier transform of the autocorrelation function yields the spectral densities or weights of the various frequencies contributing to the autocorrelation function. Using sequence and secondary structure data from more than 160 proteins and domains, a linear relationship was found between spectral density a t periodicity 3.7 and protein a-helix content ( r = 0.83). This relation permits prediction of the helix content ( x ) of proteins of known sequence to within +15%, i.e., as ( x k 15)%. Predictions based on the autocorrelation procedure are compared with values obtained by other methods. INTRODUCTIONEver since Linderstrom-Lang and Schellman' first recognized structural levels of organization within a protein, there have been many attempts to understand how protein secondary and tertiary structure is specified by the primary amino acid sequence. Some of these procedures have employed a statistical a p p r~a c h ,~-~ some a pattern-recognition a p p r~a c h ,~.~ others an information theoretic approach,'-'' and others an energy minimization approach.''?l2 One feature most of these procedures have in common is a description of the information transfer from sequence to structure in terms of the 20 commonly occurring amino acids.Am alternative approach was taken by Zimmerman et al. 13 Instead of considering structure from the point of view of the occurrence of each of the amino acids along the chain, they replaced each amino acid in a sequence with a quantitative measure of one or more of that amino acid's physical properties. Such properties were selected for their likely significance in establishing secondary and tertiary structure within the folded protein. The resulting linear series of numbers was treated as a time series and analyzed by autocorrelation methods. The essential feature of this approach is that patterns of variation of amino acid properties are observed rather than frequencies of occurrence of amino acids themselves.Jones,'* Kubota et al.,15 and most recently, Macchiato et a1.I6 have since utilized this approach, each group varying only in the properties of the amino acids included in the analysis. The original proponents, Zimmerman et al.,I3 considered that the most significant properties would be bulkiness, polarity, R , ranking in paper chromatography, acidity through the indices p1 and pK, and lastly, hydrophobicity based on solubility data of T a n f~r d . '~ Jones14 used Biopolymers, Vol. 27 451-477 (1988) 452HORNE the revised hydrophobicity scale of Nozaki and Tanford," deleted the isoionic point scale from the Zimmerman list, and included several geometric parameters of the amino acids peculiar to that particular amino acid in its particular position in an individual sequence. Kubota et al.15 described the amino acids using 10 indices, none of them geometric. The most important new additions were the propensities to form a a-helix or P-...

show abstract

Some Aspects on the Biochemistry of Growth Hormone

1988

Basic and Clinical Aspects of Growth Hormone

View full text Add to dashboard Cite

Two contiguous thrombin fragments of human somatotropin form a functionally active recombinant, but the two homologous fragments from sheep hormone do not

Cited by 18 publications

References 36 publications

Antigenic, receptor-binding and mitogenic activity of proteolytic fragments of human growth hormone.

Antigenic, receptor-binding and mitogenic activity of proteolytic fragments of human growth hormone.

Prediction of protein helix content from an autocorrelation analysis of sequence hydrophobicities

Some Aspects on the Biochemistry of Growth Hormone

Contact Info

Product

Resources

About