The Catalytic Chain of Human Complement Subcomponent C1r. Purification and N-terminal Amino Acid Sequences of the Major Cyanogen Bromide-cleavage Fragments

Overview

Journal Biochem J

Specialty Biochemistry

Date 1982 Jan 1

PMID 6282261

Citations 9

Authors

G J Arlaud

J Gagnon

R R Porter

Affiliations

Soon will be listed here.

Abstract

1. The a- and b-chains of reduced and alkylated human complement subcomponent C1r were separated by high-pressure gel-permeation chromatography and isolated in good yield and in pure form. 2. CNBr cleavage of C1r b-chain yielded eight major peptides, which were purified by gel filtration and high-pressure reversed-phase chromatography. As determined from the sum of their amino acid compositions, these peptides accounted for a minimum molecular weight of 28 000, close to the value 29 100 calculated from the whole b-chain. 3. N-Terminal sequence determinations of C1r b-chain and its CNBr-cleavage peptides allowed the identification of about two-thirds of the amino acids of C1r b-chain. From our results, and on the basis of homology with other serine proteinases, an alignment of the eight CNBr-cleavage peptides from C1r b-chain is proposed. 4. The residues forming the 'charge-relay' system of the active site of serine proteinases (His-57, Asp-102 and Ser-195 in the chymotrypsinogen numbering) are found in the corresponding regions of C1r b-chain, and the amino acid sequence around these residues has been determined. 5. The N-terminal sequence of C1r b-chain has been extended to residue 60 and reveals that C1r b-chain lacks the 'histidine loop', a disulphide bond that is present in all other known serine proteinases.

Citing Articles

Purification of human C3b inactivator by monoclonal-antibody affinity chromatography.

Hsiung L, Barclay A, Brandon M, Sim E, Porter R Biochem J. 1982; 203(1):293-8.

PMID: 7103942 PMC: 1158222. DOI: 10.1042/bj2030293.

The serine proteinase chain of human complement component C1s. Cyanogen bromide cleavage and N-terminal sequences of the fragments.

Carter P, Dunbar B, Fothergill J Biochem J. 1983; 215(3):565-71.

PMID: 6362661 PMC: 1152437. DOI: 10.1042/bj2150565.

Amino acid sequence of the Bb fragment from complement Factor B. Sequence of the major cyanogen bromide-cleavage peptide (CB-II) and completion of the sequence of the Bb fragment.

Christie D, Gagnon J Biochem J. 1983; 209(1):61-70.

PMID: 6342610 PMC: 1154056. DOI: 10.1042/bj2090061.

Autoactivation of human complement subcomponent C1r involves structural changes reflected in modifications of intrinsic fluorescence, circular dichroism and reactivity with monoclonal antibodies.

Villiers C, Arlaud G, Colomb M Biochem J. 1983; 215(2):369-75.

PMID: 6316926 PMC: 1152405. DOI: 10.1042/bj2150369.

The first component of human complement (C1): activation and control.

Ziccardi R Springer Semin Immunopathol. 1983; 6(2-3):213-30.

PMID: 6314572 DOI: 10.1007/BF00205874.

References

Woods K, Wang K . Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967; 133(2):369-70. DOI: 10.1016/0005-2795(67)90078-5. View

Naff G, Ratnoff O . The enzymatic nature of C'1r. Conversion of C'1s to C'1 esterase and digestion of amino acid esters by C'1r. J Exp Med. 1968; 128(4):571-93. PMC: 2138543. DOI: 10.1084/jem.128.4.571. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

De Bracco M, Stroud R . C1r, subunit of the first complement component: purification, properties, and assay based on its linking role. J Clin Invest. 1971; 50(4):838-48. PMC: 291998. DOI: 10.1172/JCI106555. View

Fairbanks G, Steck T, Wallach D . Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971; 10(13):2606-17. DOI: 10.1021/bi00789a030. View

Valet G, Cooper N . Isolation and characterization of the proenzyme form of the C1r subunit of the first complement component. J Immunol. 1974; 112(5):1667-73. View

Takahashi K, Nagasawa S, Koyama J . A gross structure of an activated form of a subunit of the first component of human complement. Clr. FEBS Lett. 1975; 55(1):156-60. DOI: 10.1016/0014-5793(75)80982-3. View

de Haen C, NEURATH H, Teller D . The phylogeny of trypsin-related serine proteases and their zymogens. New methods for the investigation of distant evolutionary relationships. J Mol Biol. 1975; 92(2):225-59. DOI: 10.1016/0022-2836(75)90225-9. View

Ziccardi R, Cooper N . Physicochemical and functional characterization of the C1r subunit of the first complement component. J Immunol. 1976; 116(2):496-503. View

10.

Ziccardi R, Cooper N . Activation of C1r by proteolytic cleavage. J Immunol. 1976; 116(2):504-9. View

11.

Bridgen P, Cross G, Bridgen J . N-terminal amino acid sequences of variant-specific surface antigens from Trypanosoma brucei. Nature. 1976; 263(5578):613-4. DOI: 10.1038/263613a0. View

12.

Sim R, Porter R . Isolation and comparison of the proenzyme and activated forms of the human serum complement subcomponents C1r and C1s. Biochem Soc Trans. 1976; 4(1):127-9. DOI: 10.1042/bst0040127. View

13.

Sim R, Porter R, Reid K, Gigli I . The structure and enzymic activities of the C1r and C1s subcomponents of C1, the first component of human serum complement. Biochem J. 1977; 163(2):219-27. PMC: 1164687. DOI: 10.1042/bj1630219. View

14.

Volanakis J, Schrohenloher R, Stroud R . Human factor D of the alternative complement pathway: purification and characterization. J Immunol. 1977; 119(1):337-42. View

15.

Porter R . The eleventh Hopkins Memorial Lecture: The biochemistry of complement. Biochem Soc Trans. 1977; 5(6):1659-74. DOI: 10.1042/bst0051659. View

16.

Assimeh S, Chapuis R, ISLIKER H . Studies on the precursor form of the first component of complement. II. Proteolytic fragmentation of Clr. Immunochemistry. 1978; 15(1):13-7. DOI: 10.1016/0161-5890(78)90020-2. View

17.

DODDS A, Sim R, Porter R, Kerr M . Activation of the first component of human complement (C1) by antibody-antigen aggregates. Biochem J. 1978; 175(2):383-90. PMC: 1186083. DOI: 10.1042/bj1750383. View

18.

Andrews J, Baillie R . The enzymatic nature of human c1r: a subcomponent of the first component of complement. J Immunol. 1979; 123(3):1403-8. View

19.

Arlaud G, Sim R, Duplaa A, Colomb M . Differential elution of Clq, Clr and Cls from human Cl bound to immune aggregates. Use in the rapid purification of Cl subcomponents. Mol Immunol. 1979; 16(7):445-50. DOI: 10.1016/0161-5890(79)90069-5. View

20.

Jackson C, Nemerson Y . Blood coagulation. Annu Rev Biochem. 1980; 49:765-811. DOI: 10.1146/annurev.bi.49.070180.004001. View